Formulation of diclofenac

ABSTRACT

The present invention relates to methods for producing particles of diclofenac using dry milling processes as well as compositions comprising diclofenac, medicaments produced using diclofenac in particulate form and/or compositions, and to methods of treatment of an animal, including man, using a therapeutically effective amount of diclofenac administered by way of said medicaments.

RELATED APPLICATIONS

This application is a continuation and claims priority to U.S.application Ser. No. 14/277,287, filed May 14, 2014, which is acontinuation of U.S. application Ser. No. 14/167,652, filed Jan. 29,2014, which is a continuation of U.S. application Ser. No. 13/266,122,filed Feb. 16, 2012, which is a U.S. national stage under 35 USC §371 ofInternational Application Number PCT/AU2010/000471, filed on 23 Apr.2010, which claims priority to AU Application No. 2009901748, filed on24 Apr. 2009 and U.S. Application No. 61/172,291, filed on 24 Apr. 2009,the entire contents of which applications is hereby incorporated byreference.

FIELD OF THE INVENTION

The present invention relates to methods for producing particles ofdiclofenac using dry milling processes as well as compositionscomprising diclofenac, medicaments produced using diclofenac inparticulate form and/or compositions, and to methods of treatment of ananimal, including man, using a therapeutically effective amount ofdiclofenac administered by way of said medicaments.

BACKGROUND

Poor bioavailability is a significant problem encountered in thedevelopment of compositions in the therapeutic, cosmetic, agriculturaland food industries, particularly those materials containing abiologically active material that is poorly soluble in water atphysiological pH. An active material's bioavailability is the degree towhich the active material becomes available to the target tissue in thebody or other medium after systemic administration through, for example,oral or intravenous means. Many factors affect bioavailability,including the form of dosage and the solubility and dissolution rate ofthe active material.

In therapeutic applications, poorly and slowly water-soluble materialstend to be eliminated from the gastrointestinal tract before beingabsorbed into the circulation. In addition, poorly soluble active agentstend to be disfavored or even unsafe for intravenous administration dueto the risk of particles of agent blocking blood flow throughcapillaries.

It is known that the rate of dissolution of a particulate drug willincrease with increasing surface area. One way of increasing surfacearea is decreasing particle size. Consequently, methods of making finelydivided or sized drugs have been studied with a view to controlling thesize and size range of drug particles for pharmaceutical compositions.

For example, dry milling techniques have been used to reduce particlesize and hence influence drug absorption. However, in conventional drymilling the limit of fineness is reached generally in the region ofabout 100 microns (100,000 nm), at which point material cakes on themilling chamber and prevents any further diminution of particle size.Alternatively, wet grinding may be employed to reduce particle size, butflocculation restricts the lower particle size limit to approximately 10microns (10,000 nm). The wet milling process, however, is prone tocontamination, thereby leading to a bias in the pharmaceutical artagainst wet milling. Another alternative milling technique, commercialairjet milling, has provided particles ranging in average size from aslow as about 1 to about 50 microns (1,000-50,000 nm).

There are several approaches currently used to formulate poorly solubleactive agents. One approach is to prepare the active agent as a solublesalt. Where this approach cannot be employed, alternate (usuallyphysical) approaches are employed to improve the solubility of theactive agent. Alternate approaches generally subject the active agent tophysical conditions that change the agent's physical and or chemicalproperties to improve its solubility. These include process technologiessuch as micronization, modification of crystal or polymorphic structure,development of oil based solutions, use of co-solvents, surfacestabilizers or complexing agents, micro-emulsions, supercritical fluidand production of solid dispersions or solutions. More than one of theseprocesses may be used in combination to improve formulation of aparticular therapeutic material. Many of these approaches commonlyconvert a drug into an amorphous state, which generally leads to ahigher dissolution rate. However, formulation approaches that result inthe production of amorphous material are not common in commercialformulations due to concerns relating to stability and the potential formaterial to re-crystallize.

These techniques for preparing such pharmaceutical compositions tend tobe complex. By way of example, a principal technical difficultyencountered with emulsion polymerization is the removal of contaminants,such as unreacted monomers or initiators (which may have undesirablelevels of toxicity), at the end of the manufacturing process.

Another method of providing reduced particle size is the formation ofpharmaceutical drug microcapsules, which techniques include micronizing,polymerisation and co-dispersion. However, these techniques suffer froma number of disadvantages including at least the inability to producesufficiently small particles such as those obtained by milling, and thepresence of co-solvents and/or contaminants such as toxic monomers whichare difficult to remove, leading to expensive manufacturing processes.

Over the last decade, intense scientific investigation has been carriedout to improve the solubility of active agents by converting the agentsto ultra fine powders by methods such as milling and grinding. Thesetechniques may be used to increase the dissolution rate of a particulatesolid by increasing the overall surface area and decreasing the meanparticle size.

U.S. Pat. No. 6,634,576 discloses examples of wet-milling a solidsubstrate, such as a pharmaceutically active compound, to produce a“synergetic co-mixture”.

International Patent Application PCT/AU2005/001977 (NanoparticleComposition(s) and Method for Synthesis Thereof) describes, inter alia,a method comprising the step of contacting a precursor compound with aco-reactant under mechanochemical synthesis conditions wherein asolid-state chemical reaction between the precursor compound and theco-reactant produces therapeutically active nanoparticles dispersed in acarrier matrix. Mechanochemical synthesis, as discussed in InternationalPatent Application PCT/AU2005/001977, refers to the use of mechanicalenergy to activate, initiate or promote a chemical reaction, a crystalstructure transformation or a phase change in a material or a mixture ofmaterials, for example by agitating a reaction mixture in the presenceof a milling media to transfer mechanical energy to the reactionmixture, and includes without limitation “mechanochemical activation”,“mechanochemical processing”, “reactive milling”, and related processes.

International Patent Application PCT/AU2007/000910 (Methods for thepreparation of biologically active compounds in nanoparticulate form)describes, inter alia, a method for dry milling raloxifene with lactoseand NaCl which produced nanoparticulate raloxifene without significantaggregation problems.

One limitation of many of the prior art processes is that they are notsuitable for commercial scale milling. The present invention providesmethods for overcoming the problems identified by the prior art byproviding a milling process which provides particles with increasedsurface area, yet can also be scaled up to a commercial scale.

One example of a therapeutic area where this technology could be appliedin is the area of acute pain management. Many pain medications such asdiclofenac are commonly prescribed as pain relief for chronic pain. As aresult they are commonly taken on a daily basis to maintain an effectivetherapeutic level. Diclofenac is a poorly water soluble drug sodissolution and absorption to the body is slow. So a method such as thepresent invention which provides for improved dissolution, will likelyprovide much faster absorption resulting in a more rapid onset of thetherapeutic effect. By using a method such as the present invention,which provides faster absorption, a drug such as diclofenac, could beused more readily to treat acute pain as well as chronic pain.

Although the background to the present invention is discussed in thecontext of improving the bioavailability of materials that are poorly orslowly water soluble, the applications of the methods of the presentinvention are not limited to such, as is evident from the followingdescription of the invention.

Further, although the background to the present invention is largelydiscussed in the context of improving the bioavailability of therapeuticor pharmaceutical compounds, the applications of the methods of thepresent invention are clearly not limited to such. For example, as isevident from the following description, applications of the methods ofthe present invention include but are not limited to: nutraceutical andnutritional compounds, complementary medicinal compounds, veterinarytherapeutic applications and agricultural chemical applications, such aspesticide, fungicide or herbicide.

Furthermore an application of the current invention would be tomaterials which contain a biologically active compound such as, but notlimited to a therapeutic or pharmaceutical compound, a nutraceutical ornutrient, a complementary medicinal product such as active components inplant or other naturally occurring material, a veterinary therapeuticcompound or an agricultural compound such as a pesticide, fungicide orherbicide. Specific examples would be the spice turmeric that containsthe active compound curcumin, or flax seed that contains the nutrientALA an omega 3 fatty acid. As these specific examples indicate thisinvention could be applied to, but not limited to, a range of naturalproducts such as seeds, cocoa and cocoa solids, coffee, herbs, spices,other plant materials or food materials that contain a biologicallyactive compound. The application of this invention to these types ofmaterials would enable greater availability of the active compound inthe materials when used in the relevant application. For example wherematerial subject to this invention is orally ingested the active wouldbe more bioavailable.

SUMMARY OF THE INVENTION

In one aspect the present invention is directed to the unexpectedfinding that particles of a biologically active material can be producedby dry milling processes at commercial scale. In one surprising aspectthe particle size produced by the process is equal to or less than 2000nm. In another surprising aspect the particle size produced by theprocess is equal to or less than 1000 nm. In another surprising aspectthe crystallinity of the active material is unchanged or notsubstantially changed. In a preferred embodiment the present inventionis directed to the unexpected finding that particles of diclofenac canbe produced by dry milling processes at commercial scale.

Thus in a first aspect the invention comprises a method producing acomposition, comprising the steps of dry milling a solid biologicallyactive material and a millable grinding matrix in a mill comprising aplurality of milling bodies, for a time period sufficient to produceparticles of the biologically active material dispersed in an at leastpartially milled grinding material.

In one preferred embodiment, the average particle size, determined on aparticle number basis, is equal to or less than a size selected from thegroup 2000 nm, 1900 nm, 1800 nm, 1700 nm, 1600 nm, 1500 nm, 1400 nm,1300 nm, 1200 nm, 1100 nm, 1000 nm, 900 nm, 800 nm, 700 nm, 600 nm, 500nm, 400 nm, 300 nm, 200 nm and 100 nm. Preferably, the average particlesize is equal to or greater than 25 nm.

In another preferred embodiment, the particles have a median particlesize, determined on a particle volume basis, equal or less than a sizeselected from the group 2000 nm, 1900 nm, 1800 nm, 1700 nm, 1600 nm,1500 nm, 1400 nm, 1300 nm, 1200 nm, 1100 nm, 1000 nm, 900 nm, 800 nm,700 nm, 600 nm, 500 nm, 400 nm, 300 nm, 200 nm and 100 nm. Preferably,the median particle size is equal to or greater than 25 nm. Preferably,the percentage of particles, on a particle volume basis, is selectedfrom the group consisting of: 50%, 60%, 70%, 80%, 90%, 95% and 100% lessthan 2000 nm (%<2000 nm). Preferably, the percentage of particles, on aparticle volume basis, is selected from the group consisting of: 50%,60%, 70%, 80%, 90%, 95% and 100% less than 1000 nm (%<1000 nm).Preferably, the percentage of particles, on a particle volume basis, isselected from the group consisting of: 0%, 10%, 20%, 30%, 40%, 50%, 60%,70%, 80%, 90%, 95% and 100% less than 500 nm (%<500 nm). Preferably, thepercentage of particles, on a particle volume basis, is selected fromthe group consisting of: 0%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%,90%, 95% and 100% less than 300 nm (%<300 nm). Preferably, thepercentage of particles, on a particle volume basis, is selected fromthe group consisting of: 0%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%,90%, 95% and 100% less than 200 nm (%<200 nm). Preferably, the Dx of theparticle size distribution, as measured on a particle volume basis, isselected from the group consisting of less than or equal to 10,000 nm,5000 nm, 3000 nm, 2000 nm, 1900 nm, 1800 nm, 1700 nm, 1600 nm, 1500 nm,1400 nm, 1300 nm, 1200 nm, 1100 nm, 1000 nm, 900 nm, 800 nm, 700 nm, 600nm, 500 nm, 400 nm, 300 nm, 200 nm, and 100 nm; wherein x is greaterthan or equal to 90.

In another preferred embodiment, the crystallinity profile of thebiologically active material is selected from the group consisting of:at least 50% of the biologically active material is crystalline, atleast 60% of the biologically active material is crystalline, at least70% of the biologically active material is crystalline, at least 75% ofthe biologically active material is crystalline, at least 85% of thebiologically active material is crystalline, at least 90% of thebiologically active material is crystalline, at least 95% of thebiologically active material is crystalline and at least 98% of thebiologically active material is crystalline. More preferably, thecrystallinity profile of the biologically active material issubstantially equal to the crystallinity profile of the biologicallyactive material before the material was subjected to the method asdescribed herein.

In another preferred embodiment, the amorphous content of thebiologically active material is selected from the group consisting of:less than 50% of the biologically active material is amorphous, lessthan 40% of the biologically active material is amorphous, less than 30%of the biologically active material is amorphous, less than 25% of thebiologically active material is amorphous, less than 15% of thebiologically active material is amorphous, less than 10% of thebiologically active material is amorphous, less than 5% of thebiologically active material is amorphous and less than 2% of thebiologically active material is amorphous. Preferably, the biologicallyactive material has no significant increase in amorphous content aftersubjecting the material to the method as described herein.

In another preferred embodiment, the milling time period is a rangeselected from the group consisting of: between 10 minutes and 2 hours,between 10 minutes and 90 minutes, between 10 minutes and 1 hour,between 10 minutes and 45 minutes, between 10 minutes and 30 minutes,between 5 minutes and 30 minutes, between 5 minutes and 20 minutes,between 2 minutes and 10 minutes, between 2 minutes and 5 minutes,between 1 minutes and 20 minutes, between 1 minute and 10 minutes, andbetween 1 minute and 5 minutes.

In another preferred embodiment, the milling medium is selected from thegroup consisting of: ceramics, glasses, polymers, ferromagnetics andmetals. Preferably, the milling medium is steel balls having a diameterselected from the group consisting of: between 1 and 20 mm, between 2and 15 mm and between 3 and 10 mm. In another preferred embodiment, themilling medium is zirconium oxide balls having a diameter selected fromthe group consisting of: between 1 and 20 mm, between 2 and 15 mm andbetween 3 and 10 mm. Preferably, the dry milling apparatus is a millselected from the group consisting of: attritor mills (horizontal orvertical), nutating mills, tower mills, pearl mills, planetary mills,vibratory mills, eccentric vibratory mills, gravity-dependent-type ballmills, rod mills, roller mills and crusher mills. Preferably, themilling medium within the milling apparatus is mechanically agitated by1, 2 or 3 rotating shafts. Preferably, the method is configured toproduce the biologically active material in a continuous fashion.

Preferably, the total combined amount of biologically active materialand grinding matrix in the mill at any given time is equal to or greaterthan a mass selected from the group consisting of: 200 grams, 500 grams,1 kg, 2 kg, 5 kg, 10 kg, 20 kg, 30 kg, 50 kg, 75 kg, 100 kg, 150 kg, 200kg. Preferably, the total combined amount of biologically activematerial and grinding matrix is less than 2000 kg.

Preferably, the biologically active material is selected from the groupconsisting of: diclofenac or a derivative or salt thereof.

In another preferred embodiment, the grinding matrix is a singlematerial or is a mixture of two or more materials in any proportion.Preferably, the single material or a mixture of two or more materials isselected from the group consisting of: mannitol, sorbitol, Isomalt,xylitol, maltitol, lactitol, erythritol, arabitol, ribitol, glucose,fructose, mannose, galactose, anhydrous lactose, lactose monohydrate,sucrose, maltose, trehalose, maltodextrins, dextrin, Inulin, dextrates,polydextrose, starch, wheat flour, corn flour, rice flour, rice starch,tapioca flour, tapioca starch, potato flour, potato starch, other floursand starches, milk powder, skim milk powders, other milk solids andderivatives, soy flour, soy meal or other soy products, cellulose,microcrystalline cellulose, microcrystalline cellulose based co-blendedmaterials, pregelatinized (or partially) starch, HPMC, CMC, HPC, citricacid, tartaric acid, malic acid, maleic acid fumaric acid, ascorbicacid, succinic acid, sodium citrate, sodium tartrate, sodium malate,sodium ascorbate, potassium citrate, potassium tartrate, potassiummalate, sodium acetate, potassium ascorbate, sodium carbonate, potassiumcarbonate, magnesium carbonate, sodium bicarbonate, potassiumbicarbonate, calcium carbonate, dibasic calcium phosphate, tribasiccalcium phosphate, sodium sulfate, sodium chloride, sodiummetabisulphite, sodium thiosulfate, ammonium chloride, glauber's salt,ammonium carbonate, sodium bisulfate, magnesium sulfate, potash alum,potassium chloride, sodium hydrogen sulfate, sodium hydroxide,crystalline hydroxides, hydrogen carbonates, ammonium chloride,methylamine hydrochloride, ammonium bromide, silica, thermal silica,alumina, titanium dioxide, talc, chalk, mica, kaolin, bentonite,hectorite, magnesium trisilicate, clay based materials or aluminiumsilicates, sodium lauryl sulfate, sodium stearyl sulfate, sodium cetylsulfate, sodium cetostearyl sulfate, sodium docusate, sodiumdeoxycholate, N-lauroylsarcosine sodium salt, glyceryl monostearate,glycerol distearate glyceryl palmitostearate, glyceryl behenate,glyceryl caprylate, glyceryl oleate, benzalkonium chloride, CTAB, CTAC,Cetrimide, cetylpyridinium chloride, cetylpyridinium bromide,benzethonium chloride, PEG 40 stearate, PEG 100 stearate, poloxamer 188,poloxamer 338, poloxamer 407 polyoxyl 2 stearyl ether, polyoxyl 100stearyl ether, polyoxyl 20 stearyl ether, polyoxyl 10 stearyl ether,polyoxyl 20 cetyl ether, polysorbate 20, polysorbate 40, polysorbate 60,polysorbate 61, polysorbate 65, polysorbate 80, polyoxyl 35 castor oil,polyoxyl 40 castor oil, polyoxyl 60 castor oil, polyoxyl 100 castor oil,polyoxyl 200 castor oil, polyoxyl 40 hydrogenated castor oil, polyoxyl60 hydrogenated castor oil, polyoxyl 100 hydrogenated castor oil,polyoxyl 200 hydrogenated castor oil, cetostearyl alcohol, macrogel 15hydroxystearate, sorbitan monopalmitate, sorbitan monostearate, sorbitantrioleate, sucrose palmitate, sucrose stearate, sucrose distearate,sucrose laurate, glycocholic acid, sodium glycholate, cholic acid,sodium cholate, sodium deoxycholate, deoxycholic acid, sodiumtaurocholate, taurocholic acid, sodium taurodeoxycholate,taurodeoxycholic acid, soy lecithin, phosphatidylcholine,phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol,PEG4000, PEG6000, PEG8000, PEG10000, PEG20000, alkyl naphthalenesulfonate condensate/Lignosulfonate blend, calcium dodecylbenzenesulfonate, sodium dodecylbenzene sulfonate, diisopropylnaphthaenesulphonate, erythritol distearate, naphthalene sulfonateformaldehyde condensate, nonylphenol ethoxylate (poe-30),tristyrylphenol ethoxylate, polyoxyethylene (15) tallowalkylamines,sodium alkyl naphthalene sulfonate, sodium alkyl naphthalene sulfonatecondensate, sodium alkylbenzene sulfonate, sodium isopropyl naphthalenesulfonate, sodium methyl naphthalene formaldehyde sulfonate, sodiumn-butyl naphthalene sulfonate, tridecyl alcohol ethoxylate (poe-18),triethanolamine isodecanol phosphate ester, triethanolaminetristyrylphosphate ester, tristyrylphenol ethoxylate sulfate,bis(2-hydroxyethyl)tallowalkylamines. Preferably, the concentration ofthe single (or first) material is selected from the group consisting of:5-99% w/w, 10-95% w/w, 15-85% w/w, of 20-80% w/w, 25-75% w/w, 30-60%w/w, 40-50% w/w. Preferably, the concentration of the second orsubsequent material is selected from the group consisting of: 5-50% w/w,5-40% w/w, 5-30% w/w, of 5-20% w/w, 10-40% w/w, 10-30% w/w, 10-20% w/w,20-40% w/w, or 20-30% w/w or if the second or subsequent material is asurfactant or water soluble polymer the concentration is selected from0.1-10% w/w, 0.1-5% w/w, 0.1-2.5% w/w, of 0.1-2% w/w, 0.1-1%, 0.5-5%w/w, 0.5-3% w/w, 0.5-2% w/w, 0.5-1.5%, 0.5-1% w/w, of 0.75-1.25% w/w,0.75-1% and 1% w/w.

Preferably, the grinding matrix is selected from the group consistingof:

-   -   (a) lactose monohydrate or lactose monohydrate combined with at        least one material selected from the group consisting of:        xylitol; lactose anhydrous; microcrystalline cellulose; sucrose;        glucose; sodium chloride; talc; kaolin; calcium carbonate; malic        acid; trisodium citrate dihydrate; D,L-Malic acid; sodium        pentane sulfate; sodium octadecyl sulfate; Brij700; Brij76;        sodium n-lauroyl sacrosine; lecithin; docusate sodium;        polyoxyl-40-stearate; Aerosil R972 fumed silica; sodium lauryl        sulfate or other alkyl sulfate surfactants with a chain length        between C5 to C18; polyvinyl pyrrolidone; sodium lauryl sulfate        and polyethylene glycol 40 stearate, sodium lauryl sulfate and        polyethylene glycol 100 stearate, sodium lauryl sulfate and PEG        3000, sodium lauryl sulphate and PEG 6000, sodium lauryl        sulphate and PEG 8000, sodium lauryl sulphate and PEG 10000,        sodium lauryl sulfate and Brij700, sodium lauryl sulfate and        Poloxamer 407, sodium lauryl sulfate and Poloxamer 338, sodium        lauryl sulfate and Poloxamer 188; Poloxamer 407, Poloxamer 338,        Poloxamer 188, alkyl naphthalene sulfonate        condensate/Lignosulfonate blend; Calcium Dodecylbenzene        Sulfonate (Branched); Diisopropyl naphthalenesulphonate;        erythritol distearate; linear and branched dodecylbenzene        sulfonic acids; Naphthalene Sulfonate Formaldehyde Condensate;        nonylphenol ethoxylate, POE-30; Phosphate Esters,        Tristyrylphenol Ethoxylate, Free Acid; Polyoxyethylene (15)        tallowalkylamines; sodium alkyl naphthalene sulfonate; sodium        alkyl naphthalene sulfonate condensate; sodium alkylbenzene        sulfonate; sodium isopropyl naphthalene sulfonate; Sodium Methyl        Naphthalene; Formaldehyde Sulfonate; sodium salt of n-butyl        naphthalene sulfonate; tridecyl alcohol ethoxylate, POE-18;        Triethanolamine isodecanol phosphate ester; Triethanolamine        tristyrylphosphate ester; Tristyrylphenol Ethoxylate Sulfate;        Bis(2-hydroxyethyl)tallowalkylamines.    -   (b) lactose anhydrous or lactose anhydrous combined with at        least one material selected from the group consisting of:        lactose monohydrate; xylitol; microcrystalline cellulose;        sucrose; glucose; sodium chloride; talc; kaolin; calcium        carbonate; malic acid; trisodium citrate dihydrate; D,L-Malic        acid; sodium pentane sulfate; sodium octadecyl sulfate; Brij700;        Brij76; sodium n-lauroyl sacrosine; lecithin; docusate sodium;        polyoxyl-40-stearate; Aerosil R972 fumed silica; sodium lauryl        sulfate or other alkyl sulfate surfactants with a chain length        between C5 to C18; polyvinyl pyrrolidone; sodium lauryl sulfate        and polyethylene glycol 40 stearate, sodium lauryl sulfate and        polyethylene glycol 100 stearate, sodium lauryl sulfate and PEG        3000, sodium lauryl sulphate and PEG 6000, sodium lauryl        sulphate and PEG 8000, sodium lauryl sulphate and PEG 10000,        sodium lauryl sulfate and Brij700, sodium lauryl sulfate and        Poloxamer 407, sodium lauryl sulfate and Poloxamer 338, sodium        lauryl sulfate and Poloxamer 188; Poloxamer 407, Poloxamer 338,        Poloxamer 188, alkyl naphthalene sulfonate        condensate/Lignosulfonate blend; Calcium Dodecylbenzene        Sulfonate (Branched); Diisopropyl naphthalenesulphonate;        erythritol distearate; linear and branched dodecylbenzene        sulfonic acids; Naphthalene Sulfonate Formaldehyde Condensate;        nonylphenol ethoxylate, POE-30; Phosphate Esters,        Tristyrylphenol Ethoxylate, Free Acid; Polyoxyethylene (15)        tallowalkylamines; sodium alkyl naphthalene sulfonate; sodium        alkyl naphthalene sulfonate condensate; sodium alkylbenzene        sulfonate; sodium isopropyl naphthalene sulfonate; Sodium Methyl        Naphthalene; Formaldehyde Sulfonate; sodium salt of n-butyl        naphthalene sulfonate; tridecyl alcohol ethoxylate, POE-18;        Triethanolamine isodecanol phosphate ester; Triethanolamine        tristyrylphosphate ester; Tristyrylphenol Ethoxylate Sulfate;        Bis(2-hydroxyethyl)tallowalkylamines.    -   (c) mannitol or mannitol combined with at least one material        selected from the group consisting of: lactose monohydrate;        xylitol; lactose anhydrous; microcrystalline cellulose; sucrose;        glucose; sodium chloride; talc; kaolin; calcium carbonate; malic        acid; trisodium citrate dihydrate; D,L-Malic acid; sodium        pentane sulfate; sodium octadecyl sulfate; Brij700; Brij76;        sodium n-lauroyl sacrosine; lecithin; docusate sodium;        polyoxyl-40-stearate; Aerosil R972 fumed silica; sodium lauryl        sulfate or other alkyl sulfate surfactants with a chain length        between C5 to C18; polyvinyl pyrrolidone; sodium lauryl sulfate        and polyethylene glycol 40 stearate, sodium lauryl sulfate and        polyethylene glycol 100 stearate, sodium lauryl sulfate and PEG        3000, sodium lauryl sulphate and PEG 6000, sodium lauryl        sulphate and PEG 8000, sodium lauryl sulphate and PEG 10000,        sodium lauryl sulfate and Brij700, sodium lauryl sulfate and        Poloxamer 407, sodium lauryl sulfate and Poloxamer 338, sodium        lauryl sulfate and Poloxamer 188; Poloxamer 407, Poloxamer 338,        Poloxamer 188, alkyl naphthalene sulfonate        condensate/Lignosulfonate blend; Calcium Dodecylbenzene        Sulfonate (Branched); Diisopropyl naphthalenesulphonate;        erythritol distearate; linear and branched dodecylbenzene        sulfonic acids; Naphthalene Sulfonate Formaldehyde Condensate;        nonylphenol ethoxylate, POE-30; Phosphate Esters,        Tristyrylphenol Ethoxylate, Free Acid; Polyoxyethylene (15)        tallowalkylamines; sodium alkyl naphthalene sulfonate; sodium        alkyl naphthalene sulfonate condensate; sodium alkylbenzene        sulfonate; sodium isopropyl naphthalene sulfonate; Sodium Methyl        Naphthalene; Formaldehyde Sulfonate; sodium salt of n-butyl        naphthalene sulfonate; tridecyl alcohol ethoxylate, POE-18;        Triethanolamine isodecanol phosphate ester; Triethanolamine        tristyrylphosphate ester; Tristyrylphenol Ethoxylate Sulfate;        Bis(2-hydroxyethyl)tallowalkylamines.    -   (d) Sucrose or sucrose combined with at least one material        selected from the group consisting of: lactose monohydrate;        lactose anhydrous; mannitol; microcrystalline cellulose;        glucose; sodium chloride; talc; kaolin; calcium carbonate; malic        acid; tartaric acid; trisodium citrate dihydrate; D,L-Malic        acid; sodium pentane sulfate; sodium octadecyl sulfate; Brij700;        Brij76; sodium n-lauroyl sacrosine; lecithin; docusate sodium;        polyoxyl-40-stearate; Aerosil R972 fumed silica; sodium lauryl        sulfate or other alkyl sulfate surfactants with a chain length        between C5 to C18; polyvinyl pyrrolidone; sodium lauryl sulfate        and polyethylene glycol 40 stearate, sodium lauryl sulfate and        polyethylene glycol 100 stearate, sodium lauryl sulfate and PEG        3000, sodium lauryl sulphate and PEG 6000, sodium lauryl        sulphate and PEG 8000, sodium lauryl sulphate and PEG 10000,        sodium lauryl sulfate and Brij700, sodium lauryl sulfate and        Poloxamer 407, sodium lauryl sulfate and Poloxamer 338, sodium        lauryl sulfate and Poloxamer 188; Poloxamer 407, Poloxamer 338,        Poloxamer 188, alkyl naphthalene sulfonate        condensate/Lignosulfonate blend; Calcium Dodecylbenzene        Sulfonate (Branched); Diisopropyl naphthalenesulphonate;        erythritol distearate; linear and branched dodecylbenzene        sulfonic acids; Naphthalene Sulfonate Formaldehyde Condensate;        nonylphenol ethoxylate, POE-30; Phosphate Esters,        Tristyrylphenol Ethoxylate, Free Acid; Polyoxyethylene (15)        tallowalkylamines; sodium alkyl naphthalene sulfonate; sodium        alkyl naphthalene sulfonate condensate; sodium alkylbenzene        sulfonate; sodium isopropyl naphthalene sulfonate; Sodium Methyl        Naphthalene; Formaldehyde Sulfonate; sodium salt of n-butyl        naphthalene sulfonate; tridecyl alcohol ethoxylate, POE-18;        Triethanolamine isodecanol phosphate ester; Triethanolamine        tristyrylphosphate ester; Tristyrylphenol Ethoxylate Sulfate;        Bis(2-hydroxyethyl)tallowalkylamines.    -   (e) Glucose or glucose combined with at least one material        selected from the group consisting of: lactose monohydrate;        lactose anhydrous; mannitol; microcrystalline cellulose;        sucrose; sodium chloride; talc; kaolin; calcium carbonate; malic        acid; tartaric acid; trisodium citrate dihydrate; D,L-Malic        acid; sodium pentane sulfate; sodium octadecyl sulfate; Brij700;        Brij76; sodium n-lauroyl sacrosine; lecithin; docusate sodium;        polyoxyl-40-stearate; Aerosil R972 fumed silica; sodium lauryl        sulfate or other alkyl sulfate surfactants with a chain length        between C5 to C18; polyvinyl pyrrolidone; sodium lauryl sulfate        and polyethylene glycol 40 stearate, sodium lauryl sulfate and        polyethylene glycol 100 stearate, sodium lauryl sulfate and PEG        3000, sodium lauryl sulphate and PEG 6000, sodium lauryl        sulphate and PEG 8000, sodium lauryl sulphate and PEG 10000,        sodium lauryl sulfate and Brij700, sodium lauryl sulfate and        Poloxamer 407, sodium lauryl sulfate and Poloxamer 338, sodium        lauryl sulfate and Poloxamer 188; Poloxamer 407, Poloxamer 338,        Poloxamer 188, alkyl naphthalene sulfonate        condensate/Lignosulfonate blend; Calcium Dodecylbenzene        Sulfonate (Branched); Diisopropyl naphthalenesulphonate;        erythritol distearate; linear and branched dodecylbenzene        sulfonic acids; Naphthalene Sulfonate Formaldehyde Condensate;        nonylphenol ethoxylate, POE-30; Phosphate Esters,        Tristyrylphenol Ethoxylate, Free Acid; Polyoxyethylene (15)        tallowalkylamines; sodium alkyl naphthalene sulfonate; sodium        alkyl naphthalene sulfonate condensate; sodium alkylbenzene        sulfonate; sodium isopropyl naphthalene sulfonate; Sodium Methyl        Naphthalene; Formaldehyde Sulfonate; sodium salt of n-butyl        naphthalene sulfonate; tridecyl alcohol ethoxylate, POE-18;        Triethanolamine isodecanol phosphate ester; Triethanolamine        tristyrylphosphate ester; Tristyrylphenol Ethoxylate Sulfate;        Bis(2-hydroxyethyl)tallowalkylamines.    -   (f) Sodium chloride or sodium chloride combined with at least        one material selected from the group consisting of: lactose        monohydrate; lactose anhydrous; mannitol; microcrystalline        cellulose; sucrose; glucose; talc; kaolin; calcium carbonate;        malic acid; tartaric acid; trisodium citrate dihydrate;        D,L-Malic acid; sodium pentane sulfate; sodium octadecyl        sulfate; Brij700; Brij76; sodium n-lauroyl sacrosine; lecithin;        docusate sodium; polyoxyl-40-stearate; Aerosil R972 fumed        silica; sodium lauryl sulfate or other alkyl sulfate surfactants        with a chain length between C5 to C18; polyvinyl pyrrolidone;        sodium lauryl sulfate and polyethylene glycol 40 stearate,        sodium lauryl sulfate and polyethylene glycol 100 stearate,        sodium lauryl sulfate and PEG 3000, sodium lauryl sulphate and        PEG 6000, sodium lauryl sulphate and PEG 8000, sodium lauryl        sulphate and PEG 10000, sodium lauryl sulfate and Brij700,        sodium lauryl sulfate and Poloxamer 407, sodium lauryl sulfate        and Poloxamer 338, sodium lauryl sulfate and Poloxamer 188;        Poloxamer 407, Poloxamer 338, Poloxamer 188, alkyl naphthalene        sulfonate condensate/Lignosulfonate blend; Calcium        Dodecylbenzene Sulfonate (Branched); Diisopropyl        naphthalenesulphonate; erythritol distearate; linear and        branched dodecylbenzene sulfonic acids; Naphthalene Sulfonate        Formaldehyde Condensate; nonylphenol ethoxylate, POE-30;        Phosphate Esters, Tristyrylphenol Ethoxylate, Free Acid;        Polyoxyethylene (15) tallowalkylamines; sodium alkyl naphthalene        sulfonate; sodium alkyl naphthalene sulfonate condensate; sodium        alkylbenzene sulfonate; sodium isopropyl naphthalene sulfonate;        Sodium Methyl Naphthalene; Formaldehyde Sulfonate; sodium salt        of n-butyl naphthalene sulfonate; tridecyl alcohol ethoxylate,        POE-18; Triethanolamine isodecanol phosphate ester;        Triethanolamine tristyrylphosphate ester; Tristyrylphenol        Ethoxylate Sulfate; Bis(2-hydroxyethyl)tallowalkylamines.    -   (g) xylitol or xylitol combined with at least one material        selected from the group consisting of: lactose monohydrate;        lactose anhydrous; mannitol; microcrystalline cellulose;        sucrose; glucose; sodium chloride; talc; kaolin; calcium        carbonate; malic acid; tartaric acid; trisodium citrate        dihydrate; D,L-Malic acid; sodium pentane sulfate; sodium        octadecyl sulfate; Brij700; Brij76; sodium n-lauroyl sacrosine;        lecithin; docusate sodium; polyoxyl-40-stearate; Aerosil R972        fumed silica; sodium lauryl sulfate or other alkyl sulfate        surfactants with a chain length between C5 to C18; polyvinyl        pyrrolidone; sodium lauryl sulfate and polyethylene glycol 40        stearate, sodium lauryl sulfate and polyethylene glycol 100        stearate, sodium lauryl sulfate and PEG 3000, sodium lauryl        sulphate and PEG 6000, sodium lauryl sulphate and PEG 8000,        sodium lauryl sulphate and PEG 10000, sodium lauryl sulfate and        Brij700, sodium lauryl sulfate and Poloxamer 407, sodium lauryl        sulfate and Poloxamer 338, sodium lauryl sulfate and Poloxamer        188; Poloxamer 407, Poloxamer 338, Poloxamer 188, alkyl        naphthalene sulfonate condensate/Lignosulfonate blend; Calcium        Dodecylbenzene Sulfonate (Branched); Diisopropyl        naphthalenesulphonate; erythritol distearate; linear and        branched dodecylbenzene sulfonic acids; Naphthalene Sulfonate        Formaldehyde Condensate; nonylphenol ethoxylate, POE-30;        Phosphate Esters, Tristyrylphenol Ethoxylate, Free Acid;        Polyoxyethylene (15) tallowalkylamines; sodium alkyl naphthalene        sulfonate; sodium alkyl naphthalene sulfonate condensate; sodium        alkylbenzene sulfonate; sodium isopropyl naphthalene sulfonate;        Sodium Methyl Naphthalene; Formaldehyde Sulfonate; sodium salt        of n-butyl naphthalene sulfonate; tridecyl alcohol ethoxylate,        POE-18; Triethanolamine isodecanol phosphate ester;        Triethanolamine tristyrylphosphate ester; Tristyrylphenol        Ethoxylate Sulfate; Bis(2-hydroxyethyl)tallowalkylamines.    -   (h) Tartaric acid or tartaric acid combined with at least one        material selected from the group consisting of: lactose        monohydrate; lactose anhydrous; mannitol; microcrystalline        cellulose; sucrose; glucose; sodium chloride; talc; kaolin;        calcium carbonate; malic acid; trisodium citrate dihydrate;        D,L-Malic acid; sodium pentane sulfate; sodium octadecyl        sulfate; Brij700; Brij76; sodium n-lauroyl sacrosine; lecithin;        docusate sodium; polyoxyl-40-stearate; Aerosil R972 fumed        silica; sodium lauryl sulfate or other alkyl sulfate surfactants        with a chain length between C5 to C18; polyvinyl pyrrolidone;        sodium lauryl sulfate and polyethylene glycol 40 stearate,        sodium lauryl sulfate and polyethylene glycol 100 stearate,        sodium lauryl sulfate and PEG 3000, sodium lauryl sulphate and        PEG 6000, sodium lauryl sulphate and PEG 8000, sodium lauryl        sulphate and PEG 10000, sodium lauryl sulfate and Brij700,        sodium lauryl sulfate and Poloxamer 407, sodium lauryl sulfate        and Poloxamer 338, sodium lauryl sulfate and Poloxamer 188;        Poloxamer 407, Poloxamer 338, Poloxamer 188, alkyl naphthalene        sulfonate condensate/Lignosulfonate blend; Calcium        Dodecylbenzene Sulfonate (Branched); Diisopropyl        naphthalenesulphonate; erythritol distearate; linear and        branched dodecylbenzene sulfonic acids; Naphthalene Sulfonate        Formaldehyde Condensate; nonylphenol ethoxylate, POE-30;        Phosphate Esters, Tristyrylphenol Ethoxylate, Free Acid;        Polyoxyethylene (15) tallowalkylamines; sodium alkyl naphthalene        sulfonate; sodium alkyl naphthalene sulfonate condensate; sodium        alkylbenzene sulfonate; sodium isopropyl naphthalene sulfonate;        Sodium Methyl Naphthalene; Formaldehyde Sulfonate; sodium salt        of n-butyl naphthalene sulfonate; tridecyl alcohol ethoxylate,        POE-18; Triethanolamine isodecanol phosphate ester;        Triethanolamine tristyrylphosphate ester; Tristyrylphenol        Ethoxylate Sulfate; Bis(2-hydroxyethyl)tallowalkylamines.

(i) microcrystalline cellulose or microcrystalline cellulose combinedwith at least one material selected from the group consisting of:lactose monohydrate; xylitol; lactose anhydrous; mannitol; sucrose;glucose; sodium chloride; talc; kaolin; calcium carbonate; malic acid;tartaric acid; trisodium citrate dihydrate; D,L-Malic acid; sodiumpentane sulfate; sodium octadecyl sulfate; Brij700; Brij76; sodiumn-lauroyl sacrosine; lecithin; docusate sodium; polyoxyl-40-stearate;Aerosil R972 fumed silica; sodium lauryl sulfate or other alkyl sulfatesurfactants with a chain length between C5 to C18; polyvinylpyrrolidone; sodium lauryl sulfate and polyethylene glycol 40 stearate,sodium lauryl sulfate and polyethylene glycol 100 stearate, sodiumlauryl sulfate and PEG 3000, sodium lauryl sulphate and PEG 6000, sodiumlauryl sulphate and PEG 8000, sodium lauryl sulphate and PEG 10000,sodium lauryl sulfate and Brij700, sodium lauryl sulfate and Poloxamer407, sodium lauryl sulfate and Poloxamer 338, sodium lauryl sulfate andPoloxamer 188; Poloxamer 407, Poloxamer 338, Poloxamer 188, alkylnaphthalene sulfonate condensate/Lignosulfonate blend; CalciumDodecylbenzene Sulfonate (Branched); Diisopropyl naphthalenesulphonate;erythritol distearate; linear and branched dodecylbenzene sulfonicacids; Naphthalene Sulfonate Formaldehyde Condensate; nonylphenolethoxylate, POE-30; Phosphate Esters, Tristyrylphenol Ethoxylate, FreeAcid; Polyoxyethylene (15) tallowalkylamines; sodium alkyl naphthalenesulfonate; sodium alkyl naphthalene sulfonate condensate; sodiumalkylbenzene sulfonate; sodium isopropyl naphthalene sulfonate; SodiumMethyl Naphthalene; Formaldehyde Sulfonate; sodium salt of n-butylnaphthalene sulfonate; tridecyl alcohol ethoxylate, POE-18;Triethanolamine isodecanol phosphate ester; Triethanolaminetristyrylphosphate ester; Tristyrylphenol Ethoxylate Sulfate;Bis(2-hydroxyethyl)tallowalkylamines.

(j) Kaolin combined with at least one material selected from the groupconsisting of: lactose monohydrate; xylitol; lactose anhydrous;mannitol; microcrystalline cellulose; sucrose; glucose; sodium chloride;talc; kaolin; calcium carbonate; malic acid; tartaric acid; trisodiumcitrate dihydrate; D,L-Malic acid; sodium pentane sulfate; sodiumoctadecyl sulfate; Brij700; Brij76; sodium n-lauroyl sacrosine;lecithin; docusate sodium; polyoxyl-40-stearate; Aerosil R972 fumedsilica; sodium lauryl sulfate or other alkyl sulfate surfactants with achain length between C5 to C18; polyvinyl pyrrolidone; sodium laurylsulfate and polyethylene glycol 40 stearate, sodium lauryl sulfate andpolyethylene glycol 100 stearate, sodium lauryl sulfate and PEG 3000,sodium lauryl sulphate and PEG 6000, sodium lauryl sulphate and PEG8000, sodium lauryl sulphate and PEG 10000, sodium lauryl sulfate andBrij700, sodium lauryl sulfate and Poloxamer 407, sodium lauryl sulfateand Poloxamer 338, sodium lauryl sulfate and Poloxamer 188; Poloxamer407, Poloxamer 338, Poloxamer 188, alkyl naphthalene sulfonatecondensate/Lignosulfonate blend; Calcium Dodecylbenzene Sulfonate(Branched); Diisopropyl naphthalenesulphonate; erythritol distearate;linear and branched dodecylbenzene sulfonic acids; Naphthalene SulfonateFormaldehyde Condensate; nonylphenol ethoxylate, POE-30; PhosphateEsters, Tristyrylphenol Ethoxylate, Free Acid; Polyoxyethylene (15)tallowalkylamines; sodium alkyl naphthalene sulfonate; sodium alkylnaphthalene sulfonate condensate; sodium alkylbenzene sulfonate; sodiumisopropyl naphthalene sulfonate; Sodium Methyl Naphthalene; FormaldehydeSulfonate; sodium salt of n-butyl naphthalene sulfonate; tridecylalcohol ethoxylate, POE-18; Triethanolamine isodecanol phosphate ester;Triethanolamine tristyrylphosphate ester; Tristyrylphenol EthoxylateSulfate; Bis(2-hydroxyethyl)tallowalkylamines.

-   -   (k) Talc combined with at least one material selected from the        group consisting of: lactose monohydrate; xylitol; lactose        anhydrous; mannitol; microcrystalline cellulose; sucrose;        glucose; sodium chloride; kaolin; calcium carbonate; malic acid;        tartaric acid; trisodium citrate dihydrate; D,L-Malic acid;        sodium pentane sulfate; sodium octadecyl sulfate; Brij700;        Brij76; sodium n-lauroyl sacrosine; lecithin; docusate sodium;        polyoxyl-40-stearate; Aerosil R972 fumed silica; sodium lauryl        sulfate or other alkyl sulfate surfactants with a chain length        between C5 to C18; polyvinyl pyrrolidone; sodium lauryl sulfate        and polyethylene glycol 40 stearate, sodium lauryl sulfate and        polyethylene glycol 100 stearate, sodium lauryl sulfate and PEG        3000, sodium lauryl sulphate and PEG 6000, sodium lauryl        sulphate and PEG 8000, sodium lauryl sulphate and PEG 10000,        sodium lauryl sulfate and Brij700, sodium lauryl sulfate and        Poloxamer 407, sodium lauryl sulfate and Poloxamer 338, sodium        lauryl sulfate and Poloxamer 188; Poloxamer 407, Poloxamer 338,        Poloxamer 188, alkyl naphthalene sulfonate        condensate/Lignosulfonate blend; Calcium Dodecylbenzene        Sulfonate (Branched); Diisopropyl naphthalenesulphonate;        erythritol distearate; linear and branched dodecylbenzene        sulfonic acids; Naphthalene Sulfonate Formaldehyde Condensate;        nonylphenol ethoxylate, POE-30; Phosphate Esters,        Tristyrylphenol Ethoxylate, Free Acid; Polyoxyethylene (15)        tallowalkylamines; sodium alkyl naphthalene sulfonate; sodium        alkyl naphthalene sulfonate condensate; sodium alkylbenzene        sulfonate; sodium isopropyl naphthalene sulfonate; Sodium Methyl        Naphthalene; Formaldehyde Sulfonate; sodium salt of n-butyl        naphthalene sulfonate; tridecyl alcohol ethoxylate, POE-18;        Triethanolamine isodecanol phosphate ester; Triethanolamine        tristyrylphosphate ester; Tristyrylphenol Ethoxylate Sulfate;        Bis(2-hydroxyethyl)tallowalkylamines.

Preferably, the grinding matrix is selected from the group consistingof: a material considered to be ‘Generally Regarded as Safe’ (GRAS) forpharmaceutical products; a material considered acceptable for use in anagricultural formulation; and a material considered acceptable for usein a veterinary formulation.

In another preferred embodiment, a milling aid or combination of millingaids is used. Preferably, the milling aid is selected from the groupconsisting of: colloidal silica, a surfactant, a polymer, a stearic acidand derivatives thereof. Preferably, the surfactant is selected from thegroup consisting of: polyoxyethylene alkyl ethers, polyoxyethylenestearates, polyethylene glycols (PEG), poloxamers, poloxamines,sarcosine based surfactants, polysorbates, aliphatic alcohols, alkyl andaryl sulfates, alkyl and aryl polyether sulfonates and other sulfatesurfactants, trimethyl ammonium based surfactants, lecithin and otherphospholipids, bile salts, polyoxyethylene castor oil derivatives,polyoxyethylene sorbitan fatty acid esters, Sorbitan fatty acid esters,Sucrose fatty acid esters, alkyl glucopyranosides, alkylmaltopyranosides, glycerol fatty acid esters, Alkyl Benzene SulphonicAcids, Alkyl Ether Carboxylic Acids, Alkyl and aryl Phosphate esters,Alkyl and aryl Sulphate esters, Alkyl and aryl Sulphonic acids, AlkylPhenol Phosphates esters, Alkyl Phenol Sulphates esters, Alkyl and ArylPhosphates, Alkyl Polysaccharides, Alkylamine Ethoxylates,Alkyl-Naphthalene Sulphonates formaldehyde condensates, Sulfosuccinates,lignosulfonates, Ceto-Oleyl Alcohol Ethoxylates, Condensed NaphthaleneSulphonates, Dialkyl and Alkyl Naphthalene Sulphonates, Di-alkylSulphosuccinates, Ethoxylated nonylphenols, Ethylene Glycol Esters,Fatty Alcohol Alkoxylates, Hydrogenated tallowalkylamines, Mono-alkylSulphosuccinamates, Nonyl Phenol Ethoxylates, Sodium Oleyl N-methylTaurate, Tallowalkylamines, linear and branched dodecylbenzene sulfonicacids

Preferably, the surfactant is selected from the group consisting of:sodium lauryl sulfate, sodium stearyl sulfate, sodium cetyl sulfate,sodium cetostearyl sulfate, sodium docusate, sodium deoxycholate,N-lauroylsarcosine sodium salt, glyceryl monostearate, glyceroldistearate glyceryl palmitostearate, glyceryl behenate, glycerylcaprylate, glyceryl oleate, benzalkonium chloride, CTAB, CTAC,Cetrimide, cetylpyridinium chloride, cetylpyridinium bromide,benzethonium chloride, PEG 40 stearate, PEG 100 stearate, poloxamer 188,poloxamer 338, poloxamer 407 polyoxyl 2 stearyl ether, polyoxyl 100stearyl ether, polyoxyl 20 stearyl ether, polyoxyl 10 stearyl ether,polyoxyl 20 cetyl ether, polysorbate 20, polysorbate 40, polysorbate 60,polysorbate 61, polysorbate 65, polysorbate 80, polyoxyl 35 castor oil,polyoxyl 40 castor oil, polyoxyl 60 castor oil, polyoxyl 100 castor oil,polyoxyl 200 castor oil, polyoxyl 40 hydrogenated castor oil, polyoxyl60 hydrogenated castor oil, polyoxyl 100 hydrogenated castor oil,polyoxyl 200 hydrogenated castor oil, cetostearyl alcohol, macrogel 15hydroxystearate, sorbitan monopalmitate, sorbitan monostearate, sorbitantrioleate, Sucrose Palmitate, Sucrose Stearate, Sucrose Distearate,Sucrose laurate, Glycocholic acid, sodium Glycholate, Cholic Acid,Sodium Cholate, Sodium Deoxycholate, Deoxycholic acid, Sodiumtaurocholate, taurocholic acid, Sodium taurodeoxycholate,taurodeoxycholic acid, soy lecithin, phosphatidylcholine,phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol,PEG4000, PEG6000, PEG8000, PEG10000, PEG20000, alkyl naphthalenesulfonate condensate/Lignosulfonate blend, Calcium DodecylbenzeneSulfonate, Sodium Dodecylbenzene Sulfonate, Diisopropylnaphthaenesulphonate, erythritol distearate, Naphthalene SulfonateFormaldehyde Condensate, nonylphenol ethoxylate (poe-30),Tristyrylphenol Ethoxylate, Polyoxyethylene (15) tallowalkylamines,sodium alkyl naphthalene sulfonate, sodium alkyl naphthalene sulfonatecondensate, sodium alkylbenzene sulfonate, sodium isopropyl naphthalenesulfonate, Sodium Methyl Naphthalene Formaldehyde Sulfonate, sodiumn-butyl naphthalene sulfonate, tridecyl alcohol ethoxylate (poe-18),Triethanolamine isodecanol phosphate ester, Triethanolaminetristyrylphosphate ester, Tristyrylphenol Ethoxylate Sulfate,Bis(2-hydroxyethyl)tallowalkylamines.

Preferably the polymer is selected from the list of:polyvinylpyrrolidones (PVP), polyvinylalcohol, acrylic acid basedpolymers and copolymers of acrylic acid

Preferably, the milling aid has a concentration selected from the groupconsisting of: 0.1-10% w/w, 0.1-5% w/w, 0.1-2.5% w/w, of 0.1-2% w/w,0.1-1%, 0.5-5% w/w, 0.5-3% w/w, 0.5-2% w/w, 0.5-1.5%, 0.5-1 w/w, of0.75-1.25% w/w, 0.75-1% and 1% w/w.

In another preferred embodiment of the invention, a facilitating agentis used or combination of facilitating agents is used. Preferably, thefacilitating agent is selected from the group consisting of:surfactants, polymers, binding agents, filling agents, lubricatingagents, sweeteners, flavouring agents, preservatives, buffers, wettingagents, disintegrants, effervescent agents, agents that may form part ofa medicament, including a solid dosage form or a dry powder inhalationformulation and other material required for specific drug delivery.Preferably, the facilitating agent is added during dry milling.Preferably, the facilitating agent is added to the dry milling at a timeselected from the group consisting of: with 1-5% of the total millingtime remaining, with 1-10% of the total milling time remaining, with1-20% of the total milling time remaining, with 1-30% of the totalmilling time remaining, with 2-5% of the total milling time remaining,with 2-10% of the total milling time remaining, with 5-20% of the totalmilling time remaining and with 5-20% of the total milling timeremaining. Preferably, the disintegrant is selected from the groupconsisting of: crosslinked PVP, cross linked carmellose and sodiumstarch glycolate. Preferably, the facilitating agent is added to themilled biologically active material and grinding matrix and furtherprocessed in a mechanofusion process. Mechanofusion milling causesmechanical energy to be applied to powders or mixtures of particles inthe micrometer and nanometer range.

The reasons for including facilitating agents include, but are notlimited to providing better dispersibility, control of agglomeration,the release or retention of the active particles from the deliverymatrix. Examples of facilitating agents include, but are not limited tocrosslinked PVP (crospovidone), cross linked carmellose(croscarmellose), sodium starch glycolate, Povidone (PVP), Povidone K12,Povidone K17, Povidone K25, Povidone K29/32 and Povidone K30, stearicacid, magnesium stearate, calcium stearate, sodium stearyl fumarate,sodium stearyl lactylate, zinc stearate, sodium stearate or lithiumstearate, other solid state fatty acids such as oleic acid, lauric acid,palmitic acid, erucic acid, behenic acid, or derivatives (such as estersand salts), Amino acids such as leucine, isoleucine, lysine, valine,methionine, phenylalanine, aspartame or acesulfame K. In a preferredaspect of manufacturing this formulation the facilitating agent is addedto the milled mixture of biologically active material and co-grindingmatrix and further processed in another milling device such asMechnofusion, Cyclomixing, or impact milling such as ball milling, jetmilling, or milling using a high pressure homogeniser, or combinationsthereof. In a highly preferred aspect the facilitating agent is added tothe milling of the mixture of biologically active material andco-grinding matrix as some time before the end of the milling process.

In another preferred embodiment, diclofenac is milled with lactosemonohydrate and alkyl sulfates. Preferably diclofenac is milled withlactose monohydrate and sodium lauryl sulfate. Preferably diclofenac ismilled with lactose monohydrate and sodium octadecyl sulfate. In anotherpreferred embodiment, Diclofenac is milled with lactose monohydrate,alkyl sulfates and another surfactant or polymers. Preferably diclofenacis milled with lactose monohydrate, sodium lauryl sulfate and polyethersulfates. Preferably diclofenac is milled with lactose monohydrate,sodium lauryl sulfate and polyethylene glycol 40 stearate. Preferablydiclofenac is milled with lactose monohydrate, sodium lauryl sulfate andpolyethylene glycol 100 stearate. Preferably diclofenac is milled withlactose monohydrate, sodium lauryl sulfate and a poloxamer. Preferablydiclofenac is milled with lactose monohydrate, sodium lauryl sulfate andpoloxamer 407. Preferably diclofenac is milled with lactose monohydrate,sodium lauryl sulfate and poloxamer 338. Preferably diclofenac is milledwith lactose monohydrate, sodium lauryl sulfate and poloxamer 188.Preferably diclofenac is milled with lactose monohydrate, sodium laurylsulfate and a solid polyethylene glycol. Preferably diclofenac is milledwith lactose monohydrate, sodium lauryl sulfate and polyethylene glycol6000. Preferably diclofenac is milled with lactose monohydrate, sodiumlauryl sulfate and polyethylene glycol 3000. In another preferredembodiment, Diclofenac is milled with lactose monohydrate and polyethersulfates. Preferably diclofenac is milled with lactose monohydrate andpolyethylene glycol 40 stearate. Preferably diclofenac is milled withlactose monohydrate and polyethylene glycol 100 stearate In anotherpreferred embodiment diclofenac is milled with lactose monohydrate andpolyvinyl-pyrrolidine. Preferably diclofenac is milled with lactosemonohydrate and polyvinyl-pyrrolidone with an approximate molecularweight of 30,000-40,000. In another preferred embodiment, diclofenac ismilled with lactose monohydrate and alkyl sulfonates. Preferablydiclofenac is milled with lactose monohydrate and docusate sodium. Inanother preferred embodiment, diclofenac is milled with lactosemonohydrate and a surfactant. Preferably diclofenac is milled withlactose monohydrate and lecithin. Preferably diclofenac is milled withlactose monohydrate and sodium n-lauroyl sarcosine. Preferablydiclofenac is milled with lactose monohydrate and polyoxyethylene alkylether surfactants. Preferably diclofenac is milled with lactosemonohydrate and PEG 6000. In another preferred formulation diclofenac ismilled with lactose monohydrate and silica. Preferably diclofenac ismilled with lactose monohydrate and Aerosil R972 fumed silica. Inanother preferred embodiment, diclofenac is milled with lactosemonohydrate, tartaric acid and sodium lauryl sulfate. In anotherpreferred embodiment, diclofenac is milled with lactose monohydrate,sodium bicarbonate and sodium lauryl sulfate. In another preferredembodiment, diclofenac is milled with lactose monohydrate, potassiumbicarbonate and sodium lauryl sulfate. In another preferred embodiment,diclofenac is milled with mannitol and alkyl sulfates. Preferablydiclofenac is milled with mannitol and sodium lauryl sulfate. Preferablydiclofenac is milled with mannitol and sodium octadecyl sulfate. Inanother preferred embodiment, Diclofenac is milled with mannitol, alkylsulfates and another surfactant or polymers. Preferably diclofenac ismilled with mannitol, sodium lauryl sulfate and polyether sulfates.Preferably diclofenac is milled with mannitol, sodium lauryl sulfate andpolyethylene glycol 40 stearate. Preferably diclofenac is milled withmannitol, sodium lauryl sulfate and polyethylene glycol 100 stearate.Preferably diclofenac is milled with mannitol, sodium lauryl sulfate anda poloxamer. Preferably diclofenac is milled with mannitol, sodiumlauryl sulfate and poloxamer 407. Preferably diclofenac is milled withmannitol, sodium lauryl sulfate and poloxamer 338. Preferably diclofenacis milled with mannitol, sodium lauryl sulfate and poloxamer 188.Preferably diclofenac is milled with mannitol, sodium lauryl sulfate anda solid polyethylene glycol. Preferably diclofenac is milled withmannitol, sodium lauryl sulfate and polyethylene glycol 6000. Preferablydiclofenac is milled with mannitol, sodium lauryl sulfate andpolyethylene glycol 3000. In another preferred embodiment, Diclofenac ismilled with mannitol and polyether sulfates. Preferably diclofenac ismilled with mannitol and polyethylene glycol 40 stearate. Preferablydiclofenac is milled with mannitol and polyethylene glycol 100 stearateIn another preferred embodiment diclofenac is milled with mannitol andpolyvinyl-pyrrolidine. Preferably diclofenac is milled with mannitol andpolyvinyl-pyrrolidone with an approximate molecular weight of30,000-40,000. In another preferred embodiment, diclofenac is milledwith mannitol and alkyl sulfonates. Preferably diclofenac is milled withmannitol and docusate sodium. In another preferred embodiment,diclofenac is milled with mannitol and a surfactant. Preferablydiclofenac is milled with mannitol and lecithin. Preferably diclofenacis milled with mannitol and sodium n-lauroyl sarcosine. Preferablydiclofenac is milled with mannitol and polyoxyethylene alkyl ethersurfactants. Preferably diclofenac is milled with mannitol and PEG 6000.In another preferred formulation diclofenac is milled with mannitol andsilica. Preferably diclofenac is milled with mannitol and Aerosil R972fumed silica. In another preferred embodiment, diclofenac is milled withmannitol, tartaric acid and sodium lauryl sulfate. In another preferredembodiment, diclofenac is milled with mannitol, sodium bicarbonate andsodium lauryl sulfate. In another preferred embodiment, diclofenac ismilled with mannitol, potassium bicarbonate and sodium lauryl sulfate.

In a second aspect the invention comprises a biologically activematerial produced by the method described herein and compositioncomprising the biologically active material as described herein.Preferably, the average particle size, determined on a particle numberbasis, is equal to or less than a size selected from the group 2000 nm,1900 nm, 1800 nm, 1700 nm, 1600 nm, 1500 nm, 1400 nm, 1300 nm, 1200 nm,1100 nm, 1000 nm, 900 nm, 800 nm, 700 nm, 600 nm, 500 nm, 400 nm, 300nm, 200 nm and 100 nm. Preferably, the average particle size is equal toor greater than 25 nm. Preferably, the particles have a median particlesize, determined on a particle volume basis, equal or less than a sizeselected from the group 2000 nm, 1900 nm, 1800 nm, 1700 nm, 1600 nm,1500 nm, 1400 nm, 1300 nm, 1200 nm, 1100 nm, 1000 nm, 900 nm, 800 nm,700 nm, 600 nm, 500 nm, 400 nm, 300 nm, 200 nm and 100 nm. Preferably,the median particle size is equal to or greater than 25 nm. Preferably,the percentage of particles, on a particle volume basis, is selectedfrom the group consisting of: 50%, 60%, 70%, 80%, 90%, 95% and 100% lessthan 2000 nm (%<2000 nm). Preferably, the percentage of particles, on aparticle volume basis, is selected from the group consisting of: 50%,60%, 70%, 80%, 90%, 95% and 100% less than 1000 nm (%<1000 nm).Preferably, the percentage of particles, on a particle volume basis, isselected from the group consisting of: 0%, 10%, 20%, 30%, 40%, 50%, 60%,70%, 80%, 90%, 95% and 100% less than 500 nm (%<500 nm). Preferably, thepercentage of particles, on a particle volume basis, is selected fromthe group consisting of: 0%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%,90%, 95% and 100% less than 300 nm (%<300 nm). Preferably, thepercentage of particles, on a particle volume basis, is selected fromthe group consisting of: 0%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%,90%, 95% and 100% less than 200 nm (%<200 nm). Preferably, the Dx of theparticle size distribution, as measured on a particle volume basis, isselected from the group consisting of less than or equal to 10,000 nm,5000 nm, 3000 nm, 2000 nm, 1900 nm, 1800 nm, 1700 nm, 1600 nm, 1500 nm,1400 nm, 1300 nm, 1200 nm, 1100 nm, 1000 nm, 900 nm, 800 nm, 700 nm, 600nm, 500 nm, 400 nm, 300 nm, 200 nm, and 100 nm; wherein x is greaterthan or equal to 90. Preferably, the crystallinity profile of thebiologically active material is selected from the group consisting of:at least 50% of the biologically active material is crystalline, atleast 60% of the biologically active material is crystalline, at least70% of the biologically active material is crystalline, at least 75% ofthe biologically active material is crystalline, at least 85% of thebiologically active material is crystalline, at least 90% of thebiologically active material is crystalline, at least 95% of thebiologically active material is crystalline and at least 98% of thebiologically active material is crystalline. Preferably, thecrystallinity profile of the biologically active material issubstantially equal to the crystallinity profile of the biologicallyactive material before the material was subject to the method describedherein. Preferably, the amorphous content of the biologically activematerial is selected from the group consisting of: less than 50% of thebiologically active material is amorphous, less than 40% of thebiologically active material is amorphous, less than 30% of thebiologically active material is amorphous, less than 25% of thebiologically active material is amorphous, less than 15% of thebiologically active material is amorphous, less than 10% of thebiologically active material is amorphous, less than 5% of thebiologically active material is amorphous and less than 2% of thebiologically active material is amorphous. Preferably, the biologicallyactive material has had no significant increase in amorphous contentfollowing subjecting the material to the method as described herein.

In one preferred embodiment, the invention comprises compositionscomprising the biologically active ingredient together with a grindingmatrix, a mixture of grinding matrix materials, milling aids, mixturesof milling aids, facilitating agents and/or mixtures of facilitatingagents as described herein, in concentrations and ratios as describedherein under the methods of the invention.

In a third aspect the invention comprises a pharmaceutical compositioncomprising a biologically active material produced by the methoddescribed herein and compositions described herein. Preferably, theinvention comprises pharmaceutical compositions comprising thebiologically active ingredient together with a grinding matrix, amixture of grinding matrix materials, milling aids, mixtures of millingaids, facilitating agents and/or mixtures of facilitating agents asdescribed herein, in concentrations and ratios as described herein underthe methods of the invention. Preferably, the average particle size,determined on a particle number basis, is equal to or less than a sizeselected from the group 2000 nm, 1900 nm, 1800 nm, 1700 nm, 1600 nm,1500 nm, 1400 nm, 1300 nm, 1200 nm, 1100 nm, 1000 nm, 900 nm, 800 nm,700 nm, 600 nm, 500 nm, 400 nm, 300 nm, 200 nm and 100 nm. Preferably,the average particle size is equal to or greater than 25 nm. Preferably,the particles have a median particle size, determined on a particlevolume basis, equal or less than a size selected from the group 2000 nm,1900 nm, 1800 nm, 1700 nm, 1600 nm, 1500 nm, 1400 nm, 1300 nm, 1200 nm,1100 nm, 1000 nm, 900 nm, 800 nm, 700 nm, 600 nm, 500 nm, 400 nm, 300nm, 200 nm and 100 nm. Preferably, the median particle size is equal toor greater than 25 nm. Preferably, the percentage of particles, on aparticle volume basis, is selected from the group consisting of: lessthan 2000 nm (%<2000 nm) is selected from the group consisting of: 50%,60%, 70%, 80%, 90%, 95% and 100%; less than 1000 nm (%<1000 nm) isselected from the group consisting of: 50%, 60%, 70%, 80%, 90%, 95% and100%; less than 500 nm (%<500 nm) is selected from the group 0%, 10%,20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95% and 100%; less than 300 nm(%<300 nm) is selected from the group 0%, 10%, 20%, 30%, 40%, 50%, 60%,70%, 80%, 90%, 95% and 100%; and less than 200 nm (%<200 nm) is selectedfrom the group 0%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95% and100%. Preferably, the composition has a T_(max) less than that of theequivalent conventional composition administered at the same dosage,wherein the composition comprises diclofenac. Preferably, thecomposition has a C_(max) greater than that of the equivalentconventional composition administered at the same dosage, wherein thecomposition comprises diclofenac. Preferably, the composition has an AUCgreater than that of the equivalent conventional compositionadministered at the same dosage, wherein the composition comprisesdiclofenac.

In a fourth aspect the invention comprises a method of treating a humanin need of such treatment comprising the step of administering to thehuman an effective amount of a pharmaceutical composition as describedherein.

In a fifth aspect, the invention comprises the use of a pharmaceuticalcomposition as described herein in the manufacture of a medicament forthe treatment of a human in need of such treatment.

In a sixth aspect the invention comprises a method for manufacturing apharmaceutical composition as described herein comprising the step ofcombining a therapeutically effective amount of a biologically activematerial prepared by a method described herein or a composition asdescribed herein, together with a pharmaceutically acceptable carrier toproduce a pharmaceutically acceptable dosage form.

In a seventh aspect the invention comprises a method for manufacturing aveterinary product comprising the step of combining a therapeuticallyeffective amount of the biologically active material prepared by amethod as described herein or a composition as described herein,together with an acceptable excipient to produce a dosage formacceptable for veterinary use.

In an eighth aspect the invention comprises a method for manufacturingof a pharmaceutical formulation comprising the step of combining aneffective amount of the biologically active material prepared by amethod described herein together with acceptable excipients to produce aformulation that can deliver a therapeutically effective amount ofactive to the pulmonary or nasal area. Such a formulation could be, butis not limited to a dry powder formulation for oral inhalation to thelungs or a formulation for nasal inhalation. Preferably the method formanufacturing such a formulation uses lactose, mannitol, sucrose,sorbitol, xylitol or other sugars or polyols as the co-grinding matrixtogether with surfactant such as, but not limited to lecithin, DPPC(dipalmitoyl phosphatidylcholine), PG (phosphatidylglycerol),dipalmitoyl phosphatidyl ethanolamine (DPPE), dipalmitoylphosphatidylinositol (DPPI) or other phospholipid. The particle size ofthe material produced by the invention disclosed herein results in thematerials being readily aerosolized and suitable for methods of deliveryto a subject in need thereof, including pulmonary and nasal deliverymethods.

While the method of the present invention has particular application inthe preparation of poorly water-soluble biologically active materials,the scope of the invention is not limited thereto. For example, themethod of the present invention enables production of highlywater-soluble biologically active materials. Such materials may exhibitadvantages over conventional materials by way of, for example, morerapid therapeutic action or lower dose. In contrast, wet grindingtechniques utilizing water (or other comparably polar solvents) areincapable of being applied to such materials, as the particles dissolveappreciably in the solvent.

Other aspects and advantages of the invention will become apparent tothose skilled in the art from a review of the ensuing description.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1A. Powder charge composition and particle size distribution ofmaterial milled in SPEX mill, examples A to S.

FIG. 1B. Powder charge composition and particle size distribution ofmaterial milled in SPEX mill, examples T to AL.

FIG. 1C. Powder charge composition and particle size distribution ofmaterial milled in SPEX mill, examples AM to BE.

FIG. 1D. Powder charge composition and particle size distribution ofmaterial milled in SPEX mill, examples BF to BX.

FIG. 1E. Powder charge composition and particle size distribution ofmaterial milled in SPEX mill, examples BY to CQ.

FIG. 1F. Powder charge composition and particle size distribution ofmaterial milled in SPEX mill, examples CR to DJ.

FIG. 1G. Powder charge composition and particle size distribution ofmaterial milled in SPEX mill, examples DK to EC.

FIG. 1H. The figure shows the X-Ray diffraction patterns: (A) aftermilling of Naproxen sodium in tartaric acid; (B) unmilled Naproxensodium and (C) unmilled Naproxen acid.

FIG. 2A. Powder charge composition and particle size distribution ofmaterial milled in 110 mL HD01 Attritor mill, examples A to F.

FIG. 3A. Powder charge composition and particle size distribution ofmaterial containing a mixture of 2 matrices, milled in SPEX mill,examples A to E.

FIG. 4A. Powder charge composition and particle size distribution ofmaterial milled in 1 L HD01 Attritor mill, examples A to G.

FIG. 5A. Powder charge composition and particle size distribution ofmaterial milled in 750 mL 1S Attritor mill, examples A to F.

FIG. 6A. Powder charge composition and particle size distribution ofmaterial milled in ½ Gallon 1S Attritor mill, examples A to R.

FIG. 6B. Powder charge composition and particle size distribution ofmaterial milled in ½ Gallon 1S Attritor mill, examples S to AK.

FIG. 6C. Powder charge composition and particle size distribution ofmaterial milled in ½ Gallon 1S Attritor mill, examples AL to AU.

FIG. 7A. Powder charge composition and particle size distribution ofMetaxalone milled in a variety of mills, examples A to O.

FIG. 8A. Powder charge composition and particle size distribution ofmaterial milled in HICOM mill, examples A to P.

FIG. 9A. Powder charge composition and particle size distribution ofmaterial milled in 1½ Gallon 1S Attritor mill, examples A to S.

FIG. 9B. Powder charge composition and particle size distribution ofmaterial milled in 1½ Gallon 1S Attritor mill, examples T to AL.

FIG. 10A. Powder charge composition and particle size distribution ofmaterial milled in a variety of large scale mills, examples A to F.

FIG. 11A. Powder charge composition and particle size distribution ofNaproxen Acid milled in Mannitol in a ½ Gallon 1S Attritor mill,examples A to M.

FIG. 12A. Powder charge composition and particle size distribution ofNaproxen Acid milled in SPEX mill and particle size distribution afterfiltration, examples A to L.

DETAILED DESCRIPTION OF THE INVENTION

General

Those skilled in the art will appreciate that the invention describedherein is susceptible to variations and modifications other than thosespecifically described. It is to be understood that the inventionincludes all such variations and modifications. The invention alsoincludes all of the steps, features, compositions and materials referredto or indicated in the specification, individually or collectively andany and all combinations or any two or more of the steps or features.

The present invention is not to be limited in scope by the specificembodiments described herein, which are intended for the purpose ofexemplification only. Functionally equivalent products, compositions andmethods are clearly within the scope of the invention as describedherein.

The invention described herein may include one or more ranges of values(e.g. size, concentration etc). A range of values will be understood toinclude all values within the range, including the values defining therange, and values adjacent to the range that lead to the same orsubstantially the same outcome as the values immediately adjacent tothat value which defines the boundary to the range.

The entire disclosures of all publications (including patents, patentapplications, journal articles, laboratory manuals, books, or otherdocuments) cited herein are hereby incorporated by reference. Inclusiondoes not constitute an admission is made that any of the referencesconstitute prior art or are part of the common general knowledge ofthose working in the field to which this invention relates.

Throughout this specification, unless the context requires otherwise,the word “comprise” or variations, such as “comprises” or “comprising”will be understood to imply the inclusion of a stated integer, or groupof integers, but not the exclusion of any other integers or group ofintegers. It is also noted that in this disclosure, and particularly inthe claims and/or paragraphs, terms such as “comprises”, “comprised”,“comprising” and the like can have the meaning attributed to it in USPatent law; e.g., they can mean “includes”, “included”, “including”, andthe like.

“Therapeutically effective amount” as used herein with respect tomethods of treatment and in particular drug dosage, shall mean thatdosage that provides the specific pharmacological response for which thedrug is administered in a significant number of subjects in need of suchtreatment. It is emphasized that “therapeutically effective amount,”administered to a particular subject in a particular instance will notalways be effective in treating the diseases described herein, eventhough such dosage is deemed a “therapeutically effective amount” bythose skilled in the art. It is to be further understood that drugdosages are, in particular instances, measured as oral dosages, or withreference to drug levels as measured in blood.

The term “inhibit” is defined to include its generally accepted meaningwhich includes prohibiting, preventing, restraining, and lowering,stopping, or reversing progression or severity, and such action on aresultant symptom. As such the present invention includes both medicaltherapeutic and prophylactic administration, as appropriate.

The term “biologically active material” is defined to mean abiologically active compound or a substance which comprises abiologically active compound. In this definition, a compound isgenerally taken to mean a distinct chemical entity where a chemicalformula or formulas can be used to describe the substance. Suchcompounds would generally, but not necessarily be identified in theliterature by a unique classification system such as a CAS number. Somecompounds may be more complex and have a mixed chemical structure. Forsuch compounds they may only have an empirical formula or bequalitatively identified. A compound would generally be a pure material,although it would be expected that up to 10%, 20%, 30%, 40%, 50%, 60%,70%, 80%, 90% of the substance could be other impurities and the like.Examples of biologically active compounds are, but not limited to,pharmaceutical actives, and analogs, homologs and first orderderivatives thereof. A substance that contains a biologically activecompound is any substance which has as one of its components abiologically active compound. Examples of substances containingbiologically active compounds are, but not limited to, pharmaceuticalformulations and products.

Any of the terms, “biological(ly) active”, “active”, “active material”shall have the same meaning as biologically active material.

The term “grinding matrix” is defined as any inert substance that abiologically active material can or is combined with and milled. Theterms “co-grinding matrix” and “matrix” are interchangeable with“grinding matrix”.

Particle Size

There are a wide range of techniques that can be utilized tocharacterize the particle size of a material. Those skilled in the artalso understand that almost all these techniques do not physicallymeasure the actually particle size, as one might measure something witha ruler, but measure a physical phenomena which is interpreted toindicate a particle size. As part of the interpretation process someassumptions need to be made to enable mathematical calculations to bemade. These assumptions deliver results such as an equivalent sphericalparticle size, or a hydrodynamic radius.

Amongst these various methods, two types of measurements are mostcommonly used. Photon correlation spectroscopy (PCS), also known as‘dynamic light scattering’ (DLS) is commonly used to measure particleswith a size less than 10 micron. Typically this measurement yields anequivalent hydrodynamic radius often expressed as the average size of anumber distribution. The other common particle size measurement is laserdiffraction which is commonly used to measure particle size from 100 nmto 2000 micron. This technique calculates a volume distribution ofequivalent spherical particles that can be expressed using descriptorssuch as the median particle size or the % of particles under a givensize.

Those skilled in the art recognize that different characterizationtechniques such as photon correlation spectroscopy and laser diffractionmeasure different properties of a particle ensemble. As a resultmultiple techniques will give multiple answers to the question, “what isthe particle size.” In theory one could convert and compare the variousparameters each technique measures, however, for real world particlesystems this is not practical. As a result the particle size used todescribe this invention will be given as two different sets of valuesthat each relate to these two common measurement techniques, such thatmeasurements could be made with either technique and then evaluatedagainst the description of this invention.

For measurements made using a photo correlation spectroscopy instrument,or an equivalent method known in the art, the term “number averageparticle size” is defined as the average particle diameter as determinedon a number basis.

For measurements made using a laser diffraction instrument, or anequivalent method known in the art, the term “median particle size” isdefined as the median particle diameter as determined on an equivalentspherical particle volume basis. Where the term median is used, it isunderstood to describe the particle size that divides the population inhalf such that 50% of the population is greater than or less than thissize. The median particle size is often written as D50, D(0.50) orD[0.5] or similar. As used herein D50, D(0.50) or D[0.5] or similarshall be taken to mean ‘median particle size’.

The term “Dx of the particle size distribution” refers to the xthpercentile of the distribution; thus, D90 refers to the 90^(th)percentile, D95 refers to the 95^(th) percentile, and so forth. TakingD90 as an example this can often be written as, D(0.90) or D[0.9] orsimilar. With respect to the median particle size and Dx an upper case Dor lowercase d are interchangeable and have the same meaning. Anothercommonly used way of describing a particle size distribution measured bylaser diffraction, or an equivalent method known in the art, is todescribe what % of a distribution is under or over a nominated size. Theterm “percentage less than” also written as “%<” is defined as thepercentage, by volume, of a particle size distribution under a nominatedsize—for example the %<1000 nm. The term “percentage greater than” alsowritten as “%>” is defined as the percentage, by volume, of a particlesize distribution over a nominated size—for example the %>1000 nm.

The particle size used to describe this invention should be taken tomean the particle size as measured at or shortly before the time of use.For example, the particle size is measured 2 months after the materialis subject to the milling method of this invention. In a preferred form,the particle size is measured at a time selected from the groupconsisting of: 1 day after milling, 2 days after milling, 5 days aftermilling, 1 month after milling, 2 months after milling, 3 months aftermilling, 4 months after milling, 5 months after milling, 6 months aftermilling, 1 year after milling, 2 years after milling, 5 years aftermilling.

For many of the materials subject to the methods of this invention theparticle size can be easily measured. Where the active material has poorwater solubility and the matrix it is milled in has good watersolubility the powder can simply be dispersed in an aqueous solvent. Inthis scenario the matrix dissolves leaving the active material dispersedin the solvent. This suspension can then be measured by techniques suchas PCS or laser diffraction.

Suitable methods to measure an accurate particle size where the activematerial has substantive aqueous solubility or the matrix has lowsolubility in a water based dispersant are outlined below.

-   -   1. In the circumstance where insoluble matrix such as        microcrystalline cellulose prevents the measurement of the        active material separation techniques such as filtration or        centrifugation could be used to separate the insoluble matrix        from the active material particles. Other ancillary techniques        would also be required to determine if any active material was        removed by the separation technique so that this could be taken        into account.    -   2. In the case where the active material is too soluble in water        other solvents could be evaluated for the measurement of        particle size. Where a solvent could be found that active        material is poorly soluble in but is a good solvent for the        matrix a measurement would be relatively straight forward. If        such a solvent is difficult to find another approach would be to        measure the ensemble of matrix and active material in a solvent        (such as iso-octane) which both are insoluble in. Then the        powder would be measured in another solvent where the active        material is soluble but the matrix is not. Thus with a        measurement of the matrix particle size and a measurement of the        size of the matrix and active material together an understanding        of the active material particle size can be obtained.    -   3. In some circumstances image analysis could be used to obtain        information about the particle size distribution of the active        material. Suitable image measurement techniques might include        transmission electron microscopy (TEM), scanning electron        microscopy (SEM), optical microscopy and confocal microscopy. In        addition to these standard techniques some additional technique        would be required to be used in parallel to differentiate the        active material and matrix particles. Depending on the chemical        makeup of the materials involved possible techniques could be        elemental analysis, raman spectroscopy, FTIR spectroscopy or        fluorescence spectroscopy.

OTHER DEFINITIONS

Throughout this specification, unless the context requires otherwise,the phrase “dry mill” or variations, such as “dry milling”, should beunderstood to refer to milling in at least the substantial absence ofliquids. If liquids are present, they are present in such amounts thatthe contents of the mill retain the characteristics of a dry powder.

“Flowable” means a powder having physical characteristics rendering itsuitable for further processing using typical equipment used for themanufacture of pharmaceutical compositions and formulations.

Other definitions for selected terms used herein may be found within thedetailed description of the invention and apply throughout. Unlessotherwise defined, all other scientific and technical terms used hereinhave the same meaning as commonly understood to one of ordinary skill inthe art to which the invention belongs.

The term “millable” means that the grinding matrix is capable of beingphysically degraded under the dry milling conditions of the method ofthe invention. In one embodiment of the invention, the milled grindingmatrix is of a comparable particle size to the biologically activematerial. In another embodiment of the invention the particle size ofthe matrix is substantially reduced but not as small as the biologicallyactive material

Other definitions for selected terms used herein may be found within thedetailed description of the invention and apply throughout. Unlessotherwise defined, all other scientific and technical terms used hereinhave the same meaning as commonly understood to one of ordinary skill inthe art to which the invention belongs.

Specific

In one embodiment, the present invention is directed to a method forproducing a composition, comprising the steps of: dry milling a solidbiologically active material and a millable grinding matrix in a millcomprising a plurality of milling bodies, for a time period sufficientto produce particles of the biologically active material dispersed in anat least partially milled grinding material.

The mixture of active material and matrix may then be separated from themilling bodies and removed from the mill.

In one aspect the mixture of active material and matrix is then furtherprocessed. In another aspect, the grinding matrix is separated from theparticles of biologically active material. In a further aspect, at leasta portion of the milled grinding matrix is separated from theparticulate biologically active material.

The milling bodies are essentially resistant to fracture and erosion inthe dry milling process. The quantity of the grinding matrix relative tothe quantity of biologically active material in particulate form, andthe extent of milling of the grinding matrix, is sufficient to inhibitre-agglomeration of the particles of the active material.

The present invention also relates to biologically active materialsproduced by said methods, to medicaments produced using saidbiologically active materials and to methods of treatment of an animal,including man, using a therapeutically effective amount of saidbiologically active materials administered by way of said medicaments.

Commercial Scale

The present invention is directed to the unexpected finding thatparticles of a biologically active material can be produced by drymilling processes as described herein at commercial scale. In onesurprising aspect the particle size produced by the process is equal toor less than 2000 nm. In another surprising aspect the particle sizeproduced by the process is equal to or less than 1000 nm. This canresult in a more efficient and cost effective process.

One of the key goals of reducing manufacturing costs is theencapsulation of the nanoparticles into materials that do not have to beremoved. This enables a simple manufacturing process where conventionalformulation technologies can be used to progress the matrix encapsulatednanoparticles directly to a final product. In order to do this thematerials used within the matrix must be acceptable to industryregulators. In some cases materials may be acceptable for use but onlyin limited quantities. Another aspect of matrix choice is functionality.Some matrices that produce good encapsulated nanoparticles may beacceptable from a safety perspective but these materials may makemanufacture of a dosage form such as tablet limited.

Improving the Dissolution Profile

The process results in the biologically active material having animproved dissolution profile. An improved dissolution profile hassignificant advantages including the improvement of bioavailability ofthe biologically active material in vivo. Preferably, the improveddissolution profile is observed in vitro. Alternatively, the improveddissolution profile is observed in vivo by the observation of animproved bioavailability profile. Standard methods for determining thedissolution profile of a material in vitro are available in the art. Asuitable method to determine an improved dissolution profile in vitromay include determining the concentration of the sample material in asolution over a period of time and comparing the results from the samplematerial to a control sample. An observation that peak solutionconcentration for the sample material was achieved in less time than thecontrol sample would indicate (assuming it is statisticallysignificant), that the sample material has an improved dissolutionprofile. The measurement sample is herein defined as the mixture ofbiologically active material with grinding matrix and/or other additivesthat has been subject to the processes of the invention described here.Herein a control sample is defined as a physical mixture (not subject tothe processes described in this invention) of the components in themeasurement sample with the same relative proportions of active, matrixand/or additive as the measurement sample. For the purposes of thedissolution testing a prototype formulation of the measurement samplecould also be used. In this case the control sample would be formulatedin the same way. Standard methods for determining the improveddissolution profile of a material in vivo are available in the art. Asuitable method to determine an improved dissolution profile in a humanmay be after delivering the dose to measure the rate of active materialabsorption by measuring the plasma concentration of the sample compoundover a period of time and comparing the results from the sample compoundto a control. An observation that peak plasma concentration for thesample compound was achieved in less time than the control wouldindicate (assuming it is statistically significant) that the samplecompound has improved bioavailability and an improved dissolutionprofile. Preferably, the improved dissolution profile is observed at arelevant gastrointestinal pH, when it is observed in vitro. Preferably,the improved dissolution profile is observed at a pH which is favourableat indicating improvements in dissolution when comparing the measurementsample to the control compound. Suitable methods for quantifying theconcentration of a compound in an in vitro sample or an in vivo sampleare widely available in the art. Suitable methods could include the useof spectroscopy or radioisotope labeling. In one preferred embodimentthe method of quantification of dissolution is determined in a solutionwith a pH selected from the group consisting of: pH 1, pH 2, pH 3, pH 4,pH 5, pH 6, pH 7, pH 7.3, pH 7.4, pH 8, pH 9, pH 10, pH 11, pH 12, pH13, pH 14 or a pH with 0.5 of a pH unit of any of this group.

Crystallization Profile

Methods for determining the crystallinity profile of the biologicallyactive material are widely available in the art. Suitable methods mayinclude X-ray diffraction, differential scanning calorimetry, raman orIR spectrocopy.

Amorphicity Profile

Methods for determining the amorphous content of the biologically activematerial are widely available in the art. Suitable methods may includeX-ray diffraction, differential scanning calorimetry, raman or IRspectroscopy.

Grinding Matrix

As will be described subsequently, selection of an appropriate grindingmatrix affords particular advantageous applications of the method of thepresent invention.

A highly advantageous application of the method of the invention is theuse of a water-soluble grinding matrix in conjunction with a poorlywater-soluble biologically active material. This affords at least twoadvantages. The first being when the powder containing the biologicallyactive material is placed into water—such as the ingestion of the powderas part of an oral medication—the matrix dissolves, releasing theparticulate active material such that there is maximum surface areaexposed to solution, thereby allowing a rapid dissolution of the activecompound. The second key advantage is the ability, if required, toremove or partially remove the matrix prior to further processing orformulation.

Another advantageous application of the method of the invention is theuse of a water-insoluble grinding matrix, particularly in the area ofagricultural use, when a biologically active material such as afungicide is commonly delivered as part of a dry powder or a suspension.The presence of a water insoluble matrix will afford benefits such asincreased rain fastness.

Without wishing to be bound by theory, it is believed that the physicaldegradation (including but not limited to particle size reduction) ofthe millable grinding matrix affords the advantage of the invention, byacting as a more effective diluent than grinding matrix of a largerparticle size. Again, as will be described subsequently, a highlyadvantageous aspect of the present invention is that certain grindingmatrixes appropriate for use in the method of the invention are alsoappropriate for use in a medicament. The present invention encompassesmethods for the production of a medicament incorporating both thebiologically active material and the grinding matrix or in some casesthe biologically active material and a portion of the grinding matrix,medicaments so produced, and methods of treatment of an animal,including man, using a therapeutically effective amount of saidbiologically active materials by way of said medicaments.

Analogously, as will be described subsequently, a highly advantageousaspect of the present invention is that certain grinding matrixesappropriate for use in the method of the invention are also appropriatefor use in a carrier for an agricultural chemical, such as a pesticide,fungicide, or herbicide. The present invention encompasses methods forthe production of an agricultural chemical composition incorporatingboth the biologically active material in particulate form and thegrinding matrix, or in some cases the biologically active material, anda portion of the grinding matrix, and agricultural chemical compositionsso produced. The medicament may include only the biologically activematerial together with the milled grinding matrix or, more preferably,the biologically active material and milled grinding matrix may becombined with one or more pharmaceutically acceptable carriers, as wellas any desired excipients or other like agents commonly used in thepreparation of medicaments.

Analogously, the agricultural chemical composition may include only thebiologically active material together with the milled grinding matrixor, more preferably, the biologically active materials and milledgrinding matrix may be combined with one or more carriers, as well asany desired excipients or other like agents commonly used in thepreparation of agricultural chemical compositions.

In one particular form of the invention, the grinding matrix is bothappropriate for use in a medicament and readily separable from thebiologically active material by methods not dependent on particle size.Such grinding matrixes are described in the following detaileddescription of the invention. Such grinding matrixes are highlyadvantageous in that they afford significant flexibility in the extentto which the grinding matrix may be incorporated with the biologicallyactive material into a medicament.

In a highly preferred form, the grinding matrix is harder than thebiologically active material, and is thus capable of reducing theparticle size of the active material under the dry milling conditions ofthe invention. Again without wishing to be bound by theory, under thesecircumstances it is believed that the millable grinding matrix affordsthe advantage of the present invention through a second route, with thesmaller particles of grinding matrix produced under the dry millingconditions enabling greater interaction with the biologically activematerial. The quantity of the grinding matrix relative to the quantityof biologically active material, and the extent of physical degradationof the grinding matrix, is sufficient to inhibit re-agglomeration of theparticles of the active material Preferably, the quantity of thegrinding matrix relative to the quantity of biologically activematerial, and the extent of physical degradation of the grinding matrix,is sufficient to inhibit re-agglomeration of the particles of the activematerial in nanoparticulate form. The grinding matrix is not generallyselected to be chemically reactive with the biologically active materialunder the milling conditions of the invention, excepting for example,where the matrix is deliberately chosen to undergo a mechanico-chemicalreaction. Such a reaction might be the conversion of a free base or acidto a salt or vice versa.

As stated above, the method of the present invention requires thegrinding matrix to be milled with the biologically active material; thatis, the grinding matrix will physically degrade under the dry millingconditions of the invention to facilitate the formation and retention ofparticulates of the biologically active material with reduced particlesize. The precise extent of degradation required will depend on certainproperties of the grinding matrix and the biologically active material,the ratio of biologically active material to grinding matrix, and theparticle size distribution of the particles comprising the biologicallyactive material.

The physical properties of the grinding matrix necessary to achieve therequisite degradation are dependent on the precise milling conditions.For example, a harder grinding matrix may degrade to a sufficient extentprovided it is subjected to more vigorous dry milling conditions.

Physical properties of the grinding matrix relevant to the extent thatthe agent will degrade under dry milling conditions include hardness,friability, as measured by indicia such as hardness, fracture toughnessand brittleness index.

A low hardness (typically a Mohs Hardness less than 7) of thebiologically active material is desirable to ensure fracture of theparticles during processing, so that composite microstructures developduring milling. Preferably, the hardness is less than 3 as determinedusing the Mohs Hardness scale.

Preferably, the grinding matrix is of low abrasivity. Low abrasivity isdesirable to minimise contamination of the mixture of the biologicallyactive material in the grinding matrix by the milling bodies and/or themilling chamber of the media mill. An indirect indication of theabrasivity can be obtained by measuring the level of milling-basedcontaminants.

Preferably, the grinding matrix has a low tendency to agglomerate duringdry milling. While it is difficult to objectively quantify the tendencyto agglomerate during milling, it is possible to obtain a subjectivemeasure by observing the level of “caking” of the grinding matrix on themilling bodies and the milling chamber of the media mill as dry millingprogresses.

The grinding matrix may be an inorganic or organic substance.

In one embodiment, the grinding matrix is selected from the following,either as a single substance or a combination of two or more substances:Polyols (sugar alcohols) for example (but not limited to) mannitol,sorbitol, isomalt, xylitol, maltitol, lactitol, erythritol, arabitol,ribitol, monosaccharides for example (but not limited to) glucose,fructose, mannose, galactose, disaccharides and trisaccharides forexample (but not limited to) anhydrous lactose, lactose monohydrate,sucrose, maltose, trehalose, polysaccharides for example (but notlimited to) maltodextrins, dextrin, Inulin, dextrates, polydextrose,other carbohydrates for example (but not limited to) starch, wheatflour, corn flour, rice flour, rice starch, tapioca flour, tapiocastarch, potato flour, potato starch, other flours and starches, soyflour, soy meal or other soy products, cellulose, microcrystallinecellulose, microcrystalline cellulose based co blended excipients,chemically modified excipients such as pregelatinized (or partially)starch, modified celluloses such as HPMC, CMC, HPC, enteric polymercoatings such as hypromellose phthalate, cellulose acetate phthalate(Aquacoat®), polyvinyl acetate phthalate (Sureteric®), hypromelloseacetate succinate (AQOAT®), and polmethacrylates (Eudragit® andAcryl-EZE®), Milk products for example (but not limited to) milk powder,skim milk powders, other milk solids and derivatives, other functionalExcipients, organic acids for example (but not limited to) citric acid,tartaric acid, malic acid, maleic acid fumaric acid, ascorbic acid,succinic acid, the conjugate salt of organic acids for example (but notlimited to) sodium citrate, sodium tartrate, sodium malate, sodiumascorbate, potassium citrate, potassium tartrate, potassium malate,potassium ascorbate, inorganics such as sodium carbonate, potassiumcarbonate, magnesium carbonate, sodium bicarbonate, potassiumbicarbonate and calcium carbonate. dibasic calcium phosphate, tribasiccalcium phosphate, sodium sulfate, sodium chloride, sodiummetabisulphite, sodium thiosulfate, ammonium chloride, Glauber's salt,ammonium carbonate, sodium bisulfate, magnesium sulfate, potash alum,potassium chloride, sodium hydrogen sulfate, sodium hydroxide,crystalline hydroxides, hydrogen carbonates, hydrogen carbonates ofpharmaceutical acceptable alkali metals, such as but not limited by,sodium, potassium, lithium, calcium, and barium, ammonium salts (orsalts of volatile amines), for example (but not limited to) ammoniumchloride, methylamine hydrochloride, ammonium bromide, other inorganicsfor example (but not limited to), thermal silica, chalk, mica, silica,alumina, titanium dioxide, talc, kaolin, bentonite, hectorite, magnesiumtrisilicate, other clay or clay derivatives or aluminium silicates, asurfactant for example (but not limited to) sodium lauryl sulfate,sodium stearyl sulfate, sodium cetyl sulfate, sodium cetostearylsulfate, sodium docusate, sodium deoxycholate, N-lauroylsarcosine sodiumsalt, glyceryl monostearate, glycerol distearate glycerylpalmitostearate, glyceryl behenate, glyceryl caprylate, glyceryl oleate,benzalkonium chloride, CTAB, CTAC, Cetrimide, cetylpyridinium chloride,cetylpyridinium bromide, benzethonium chloride, PEG 40 stearate, PEG 100stearate, poloxamer 188, poloxamer 407, poloxamer 338, polyoxyl 2stearyl ether, polyoxyl 100 stearyl ether, polyoxyl 20 stearyl ether,polyoxyl 10 stearyl ether, polyoxyl 20 cetyl ether, polysorbate 20,polysorbate 40, polysorbate 60, polysorbate 61, polysorbate 65,polysorbate 80, polyoxyl 35 castor oil, polyoxyl 40 castor oil, polyoxyl60 castor oil, polyoxyl 100 castor oil, polyoxyl 200 castor oil,polyoxyl 40 hydrogenated castor oil, polyoxyl 60 hydrogenated castoroil, polyoxyl 100 hydrogenated castor oil, polyoxyl 200 hydrogenatedcastor oil, cetostearyl alcohol, macrogel 15 hydroxystearate, sorbitanmonopalmitate, sorbitan monostearate, sorbitan trioleate, SucrosePalmitate, Sucrose Stearate, Sucrose Distearate, Sucrose laurate,Glycocholic acid, sodium Glycholate, Cholic Acid, Sodium Cholate, SodiumDeoxycholate, Deoxycholic acid, Sodium taurocholate, taurocholic acid,Sodium taurodeoxycholate, taurodeoxycholic acid, soy lecithin,phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine,phosphatidylinositol, PEG4000, PEG6000, PEG8000, PEG10000, PEG20000,alkyl naphthalene sulfonate condensate/Lignosulfonate blend, CalciumDodecylbenzene Sulfonate, Sodium Dodecylbenzene Sulfonate, Diisopropylnaphthaenesulphonate, erythritol distearate, Naphthalene SulfonateFormaldehyde Condensate, nonylphenol ethoxylate (poe-30),Tristyrylphenol Ethoxylate, Polyoxyethylene (15) tallowalkylamines,sodium alkyl naphthalene sulfonate, sodium alkyl naphthalene sulfonatecondensate, sodium alkylbenzene sulfonate, sodium isopropyl naphthalenesulfonate, Sodium Methyl Naphthalene Formaldehyde Sulfonate, sodiumn-butyl naphthalene sulfonate, tridecyl alcohol ethoxylate (poe-18),Triethanolamine isodecanol phosphate ester, Triethanolaminetristyrylphosphate ester, Tristyrylphenol Ethoxylate Sulfate,Bis(2-hydroxyethyl)tallowalkylamines.

In a preferred embodiment, the grinding matrix is a matrix that isconsidered GRAS (generally regarded as safe) by persons skilled in thepharmaceutical arts.

In another preferred aspect a combination of two or more suitablematrices, such as those listed above, can be used as the grinding matrixto provide improved properties such as the reduction of caking, andgreater improvement of the dissolution profile. Combination matrices mayalso be advantageous when the matrices have different solubility'sallowing the removal or partial removal of one matrix, while leaving theother or part of the other to provide encapsulation or partialencapsulation of the biologically active material.

Another highly preferred aspect of the method is the inclusion of asuitable milling aid in the matrix to improve milling performance.Improvements to milling performance would be things such as, but notlimited to, a reduction in caking or higher recovery of powder from themill. Examples of suitable milling aids include surfactants, polymersand inorganics such as silica (including colloidal silica), aluminiumsilicates and clays.

There are a wide range of surfactants that will make suitable millingaids. The highly preferred form is where the surfactant is a solid, orcan be manufactured into a solid. Preferably, the surfactant is selectedfrom the group consisting of: polyoxyethylene alkyl ethers,polyoxyethylene stearates, polyethylene glycols (PEG), poloxamers,poloxamines, sarcosine based surfactants, polysorbates, aliphaticalcohols, alkyl and aryl sulfates, alkyl and aryl polyether sulfonatesand other sulfate surfactants, trimethyl ammonium based surfactants,lecithin and other phospholipids, bile salts, polyoxyethylene castor oilderivatives, polyoxyethylene sorbitan fatty acid esters, Sorbitan fattyacid esters, Sucrose fatty acid esters, alkyl glucopyranosides, alkylmaltopyranosides, glycerol fatty acid esters, Alkyl Benzene SulphonicAcids, Alkyl Ether Carboxylic Acids, Alkyl and aryl Phosphate esters,Alkyl and aryl Sulphate esters, Alkyl and aryl Sulphonic acids, AlkylPhenol Phosphates esters, Alkyl Phenol Sulphates esters, Alkyl and ArylPhosphates, Alkyl Polysaccharides, Alkylamine Ethoxylates,Alkyl-Naphthalene Sulphonates formaldehyde condensates, Sulfosuccinates,lignosulfonates, Ceto-Oleyl Alcohol Ethoxylates, Condensed NaphthaleneSulphonates, Dialkyl and Alkyl Naphthalene Sulphonates, Di-alkylSulphosuccinates, Ethoxylated nonylphenols, Ethylene Glycol Esters,Fatty Alcohol Alkoxylates, Hydrogenated tallowalkylamines, Mono-alkylSulphosuccinamates, Nonyl Phenol Ethoxylates, Sodium Oleyl N-methylTaurate, Tallowalkylamines, linear and branched dodecylbenzene sulfonicacids Preferably, the surfactant is selected from the group consistingof: sodium lauryl sulfate, sodium stearyl sulfate, sodium cetyl sulfate,sodium cetostearyl sulfate, sodium docusate, sodium deoxycholate,N-lauroylsarcosine sodium salt, glyceryl monostearate, glyceroldistearate glyceryl palmitostearate, glyceryl behenate, glycerylcaprylate, glyceryl oleate, benzalkonium chloride, CTAB, CTAC,Cetrimide, cetylpyridinium chloride, cetylpyridinium bromide,benzethonium chloride, PEG 40 stearate, PEG 100 stearate, poloxamer 188,poloxamer 338, poloxamer 407 polyoxyl 2 stearyl ether, polyoxyl 100stearyl ether, polyoxyl 20 stearyl ether, polyoxyl 10 stearyl ether,polyoxyl 20 cetyl ether, polysorbate 20, polysorbate 40, polysorbate 60,polysorbate 61, polysorbate 65, polysorbate 80, polyoxyl 35 castor oil,polyoxyl 40 castor oil, polyoxyl 60 castor oil, polyoxyl 100 castor oil,polyoxyl 200 castor oil, polyoxyl 40 hydrogenated castor oil, polyoxyl60 hydrogenated castor oil, polyoxyl 100 hydrogenated castor oil,polyoxyl 200 hydrogenated castor oil, cetostearyl alcohol, macrogel 15hydroxystearate, sorbitan monopalmitate, sorbitan monostearate, sorbitantrioleate, Sucrose Palmitate, Sucrose Stearate, Sucrose Distearate,Sucrose laurate, Glycocholic acid, sodium Glycholate, Cholic Acid,Sodium Cholate, Sodium Deoxycholate, Deoxycholic acid, Sodiumtaurocholate, taurocholic acid, Sodium taurodeoxycholate,taurodeoxycholic acid, soy lecithin, phosphatidylcholine,phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol,PEG4000, PEG6000, PEG8000, PEG10000, PEG20000, alkyl naphthalenesulfonate condensate/Lignosulfonate blend, Calcium DodecylbenzeneSulfonate, Sodium Dodecylbenzene Sulfonate, Diisopropylnaphthaenesulphonate, erythritol distearate, Naphthalene SulfonateFormaldehyde Condensate, nonylphenol ethoxylate (poe-30),Tristyrylphenol Ethoxylate, Polyoxyethylene (15) tallowalkylamines,sodium alkyl naphthalene sulfonate, sodium alkyl naphthalene sulfonatecondensate, sodium alkylbenzene sulfonate, sodium isopropyl naphthalenesulfonate, Sodium Methyl Naphthalene Formaldehyde Sulfonate, sodiumn-butyl naphthalene sulfonate, tridecyl alcohol ethoxylate (poe-18),Triethanolamine isodecanol phosphate ester, Triethanolaminetristyrylphosphate ester, Tristyrylphenol Ethoxylate Sulfate,Bis(2-hydroxyethyl)tallowalkylamines.

Preferably the polymer is selected from the list of:polyvinylpyrrolidones (PVP), polyvinylalcohol, Acrylic acid basedpolymers and copolymers of acrylic acid

Preferably, the milling aid has a concentration selected from the groupconsisting of: 0.1-10% w/w, 0.1-5% w/w, 0.1-2.5% w/w, of 0.1-2% w/w,0.1-1%, 0.5-5% w/w, 0.5-3% w/w, 0.5-2% w/w, 0.5-1.5%, 0.5-1 w/w, of0.75-1.25% w/w, 0.75-1% and 1% w/w.

Milling Bodies

In the method of the present invention, the milling bodies arepreferably chemically inert and rigid. The term “chemically-inert”, asused herein, means that the milling bodies do not react chemically withthe biologically active material or the grinding matrix.

As described above, the milling bodies are essentially resistant tofracture and erosion in the milling process.

The milling bodies are desirably provided in the form of bodies whichmay have any of a variety of smooth, regular shapes, flat or curvedsurfaces, and lacking sharp or raised edges. For example, suitablemilling bodies can be in the form of bodies having ellipsoidal, ovoid,spherical or right cylindrical shapes. Preferably, the milling bodiesare provided in the form of one or more of beads, balls, spheres, rods,right cylinders, drums or radius-end right cylinders (i.e., rightcylinders having hemispherical bases with the same radius as thecylinder).

Depending on the nature of the biologically active material and thegrinding matrix, the milling media bodies desirably have an effectivemean particle diameter (i.e. “particle size”) between about 0.1 and 30mm, more preferably between about 1 and about 15 mm, still morepreferably between about 3 and 10 mm.

The milling bodies may comprise various substances such as ceramic,glass, metal or polymeric compositions, in a particulate form. Suitablemetal milling bodies are typically spherical and generally have goodhardness (i.e. RHC 60-70), roundness, high wear resistance, and narrowsize distribution and can include, for example, balls fabricated fromtype 52100 chrome steel, type 316 or 440C stainless steel or type 1065high carbon steel.

Preferred ceramics, for example, can be selected from a wide array ofceramics desirably having sufficient hardness and resistance to fractureto enable them to avoid being chipped or crushed during milling and alsohaving sufficiently high density. Suitable densities for milling mediacan range from about 1 to 15 g/cm³′, preferably from about 1 to 8 g/cm³.Preferred ceramics can be selected from steatite, aluminum oxide,zirconium oxide, zirconia-silica, yttria-stabilized zirconium oxide,magnesia-stabilized zirconium oxide, silicon nitride, silicon carbide,cobalt-stabilized tungsten carbide, and the like, as well as mixturesthereof.

Preferred glass milling media are spherical (e.g. beads), have a narrowsize distribution, are durable, and include, for example, lead-free sodalime glass and borosilicate glass. Polymeric milling media arepreferably substantially spherical and can be selected from a wide arrayof polymeric resins having sufficient hardness and friability to enablethem to avoid being chipped or crushed during milling,abrasion-resistance to minimize attrition resulting in contamination ofthe product, and freedom from impurities such as metals, solvents, andresidual monomers.

Preferred polymeric resins, for example, can be selected fromcrosslinked polystyrenes, such as polystyrene crosslinked withdivinylbenzene, styrene copolymers, polyacrylates such aspolymethylmethacrylate, polycarbonates, polyacetals, vinyl chloridepolymers and copolymers, polyurethanes, polyamides, high densitypolyethylenes, polypropylenes, and the like. The use of polymericmilling media to grind materials down to a very small particle size (asopposed to mechanochemical synthesis) is disclosed, for example, in U.S.Pat. Nos. 5,478,705 and 5,500,331. Polymeric resins typically can havedensities ranging from about 0.8 to 3.0 g/cm³. Higher density polymericresins are preferred. Alternatively, the milling media can be compositeparticles comprising dense core particles having a polymeric resinadhered thereon. Core particles can be selected from substances known tobe useful as milling media, for example, glass, alumina, zirconiasilica, zirconium oxide, stainless steel, and the like. Preferred coresubstances have densities greater than about 2.5 g/cm³.

In one embodiment of the invention, the milling media are formed from aferromagnetic substance, thereby facilitating removal of contaminantsarising from wear of the milling media by the use of magnetic separationtechniques.

Each type of milling body has its own advantages. For example, metalshave the highest specific gravities, which increase grinding efficiencydue to increased impact energy. Metal costs range from low to high, butmetal contamination of final product can be an issue. Glasses areadvantageous from the standpoint of low cost and the availability ofsmall bead sizes as low as 0.004 mm. However, the specific gravity ofglasses is lower than other media and significantly more milling time isrequired. Finally, ceramics are advantageous from the standpoint of lowwear and contamination, ease of cleaning, and high hardness.

Dry Milling

In the dry milling process of the present invention, the biologicallyactive material and grinding matrix, in the form of crystals, powders,or the like, are combined in suitable proportions with the plurality ofmilling bodies in a milling chamber that is mechanically agitated (i.e.with or without stirring) for a predetermined period of time at apredetermined intensity of agitation. Typically, a milling apparatus isused to impart motion to the milling bodies by the external applicationof agitation, whereby various translational, rotational or inversionmotions or combinations thereof are applied to the milling chamber andits contents, or by the internal application of agitation through arotating shaft terminating in a blade, propeller, impeller or paddle orby a combination of both actions.

During milling, motion imparted to the milling bodies can result inapplication of shearing forces as well as multiple impacts or collisionshaving significant intensity between milling bodies and particles of thebiologically active material and grinding matrix. The nature andintensity of the forces applied by the milling bodies to thebiologically active material and the grinding matrix is influenced by awide variety of processing parameters including: the type of millingapparatus; the intensity of the forces generated, the kinematic aspectsof the process; the size, density, shape, and composition of the millingbodies; the weight ratio of the biologically active material andgrinding matrix mixture to the milling bodies; the duration of milling;the physical properties of both the biologically active material and thegrinding matrix; the atmosphere present during activation; and others.

Advantageously, the media mill is capable of repeatedly or continuouslyapplying mechanical compressive forces and shear stress to thebiologically active material and the grinding matrix. Suitable mediamills include but are not limited to the following: high-energy ball,sand, bead or pearl mills, basket mill, planetary mill, vibratory actionball mill, multi-axial shaker/mixer, stirred ball mill, horizontal smallmedia mill, multi-ring pulverizing mill, and the like, including smallmilling media. The milling apparatus also can contain one or morerotating shafts.

In a preferred form of the invention, the dry milling is performed in aball mill. Throughout the remainder of the specification reference willbe made to dry milling being carried out by way of a ball mill. Examplesof this type of mill are attritor mills, nutating mills, tower mills,planetary mills, vibratory mills and gravity-dependent-type ball mills.It will be appreciated that dry milling in accordance with the method ofthe invention may also be achieved by any suitable means other than ballmilling. For example, dry milling may also be achieved using jet mills,rod mills, roller mills or crusher mills.

Biologically Active Material

The biologically active material includes active compounds, includingcompounds for veterinary and human use such as but not limited to,pharmaceutical actives and the like.

The biologically active material is ordinarily a material for which oneof skill in the art desires improved dissolution properties. Thebiologically active material may be a conventional active agent or drug,although the process of the invention may be employed on formulations oragents that already have reduced particle size compared to theirconventional form.

Biologically active materials suitable for use in the invention includediclofenac.

As discussed in the context of the background to the invention,biologically active materials that are poorly water soluble atgastrointestinal pH will particularly benefit from being prepared, andthe method of the present invention is particularly advantageouslyapplied to materials that are poorly water soluble at gastrointestinalpH.

Conveniently, the biologically active material is capable ofwithstanding temperatures that are typical in uncooled dry milling,which may exceed 80° C. Therefore, materials with a melting point about80° C. or greater are highly suitable. For biologically active materialswith lower melting points, the media mill may be cooled, therebyallowing materials with significantly lower melting temperatures to beprocessed according to the method of the invention. For instance, asimple water-cooled mill will keep temperatures below 50° C., or chilledwater could be used to further lower the milling temperature. Thoseskilled in the art will understand that a high energy ball mill could bedesigned to run at any temperature between say −30 to 200° C. For somebiologically active materials it may be advantageous to control themilling temperature to temperatures significantly below the meltingpoints of the biologically active materials.

The biologically active material is obtained in a conventional formcommercially and/or prepared by techniques known in the art.

It is preferred, but not essential, that the particle size of thebiologically active material be less than about 1000 μm, as determinedby sieve analysis. If the coarse particle size of the biologicallyactive material is greater than about 1000 μm, then it is preferred thatthe particles of the biologically active material substrate be reducedin size to less than 1000 μm using another standard milling method.

Processed Biologically Active Material

Preferably, the biologically active materials, which have been subjectto the methods of the invention, comprises particles of biologicallyactive material of an average particle size, determined on a particlenumber basis, is equal to or less than a size selected from the group2000 nm, 1900 nm, 1800 nm, 1700 nm, 1600 nm, 1500 nm, 1400 nm, 1300 nm,1200 nm, 1100 nm, 1000 nm, 900 nm, 800 nm, 700 nm, 600 nm, 500 nm, 400nm, 300 nm, 200 nm and 100 nm. Preferably, the biologically activematerials, which have been subject to the methods of the invention,comprises particles of biologically active material of a median particlesize, determined on a particle volume basis, equal or less than a sizeselected from the group 2000 nm, 1900 nm, 1800 nm, 1700 nm, 1600 nm,1500 nm, 1400 nm, 1300 nm, 1200 nm, 1100 nm, 1000 nm, 900 nm, 800 nm,700 nm, 600 nm, 500 nm, 400 nm, 300 nm, 200 nm and 100 nm.

Preferably, the biologically active materials, which have been subjectto the methods of the invention, comprises particles of biologicallyactive material and wherein the Dx of the particle size distribution, asmeasured on a particle volume basis, is selected from the groupconsisting of less than or equal to 10,000 nm, 5000 nm, 3000 nm, 2000nm, 1900 nm, 1800 nm, 1700 nm, 1600 nm, 1500 nm, 1400 nm, 1300 nm, 1200nm, 1100 nm, 1000 nm, 900 nm, 800 nm, 700 nm, 600 nm, 500 nm, 400 nm,300 nm, 200 nm, and 100 nm; wherein x is greater than or equal to 90,These sizes refer to particles either fully dispersed or partiallyagglomerated.

Agglomerates of Biologically Active Material after Processing

Agglomerates comprising particles of biologically active material, saidparticles having a particle size within the ranges specified above,should be understood to fall within the scope of the present invention,regardless of whether the agglomerates exceed the ranges specifiedabove.

Agglomerates comprising particles of biologically active material, saidagglomerates having a total agglomerate size within the ranges specifiedabove, should be understood to fall within the scope of the presentinvention.

Agglomerates comprising particles of biologically active material shouldbe understood to fall within the scope of the present invention if atthe time of use, or further processing, the particle size of theagglomerate is within the ranges specified above.

Agglomerates comprising particles of biologically active material, saidparticles having a particle size within the ranges specified above, atthe time of use, or further processing, should be understood to fallwithin the scope of the present invention, regardless of whether theagglomerates exceed the ranges specified above.

Processing Time

Preferably, the biologically active material and the grinding matrix aredry milled for the shortest time necessary to form the mixture of thebiologically active material in the grinding matrix such that the activematerial has improved dissolution to minimise any possible contaminationfrom the media mill and/or the plurality of milling bodies. This timevaries greatly, depending on the biologically active material and thegrinding matrix, and may range from as short as 1 minute to severalhours. Dry milling times in excess of 2 hours may lead to degradation ofthe biologically active material and an increased level of undesirablecontaminants.

Suitable rates of agitation and total milling times are adjusted for thetype and size of milling apparatus as well as the milling media, theweight ratio of the biologically active material and grinding matrixmixture to the plurality of milling bodies, the chemical and physicalproperties of the biologically active material and grinding matrix, andother parameters that may be optimized empirically.

Inclusion of the Grinding Matrix with the Biologically Active Materialand Separation of the Grinding Matrix from the Biologically ActiveMaterial

In a preferred aspect, the grinding matrix is not separated from thebiologically active material but is maintained with the biologicallyactive material in the final product. Preferably the grinding matrix isconsidered to be Generally Regarded as Safe (GRAS) for pharmaceuticalproducts.

In an alternative aspect, the grinding matrix is separated from thebiologically active material. In one aspect, where the grinding matrixis not fully milled, the unmilled grinding matrix is separated from thebiologically active material. In a further aspect, at least a portion ofthe milled grinding matrix is separated from the biologically activematerial.

Any portion of the grinding matrix may be removed, including but notlimited to 10%, 25%, 50%, 75%, or substantially all of the grindingmatrix.

In some embodiments of the invention, a significant portion of themilled grinding matrix may comprise particles of a size similar toand/or smaller than the particles comprising the biologically activematerial. Where the portion of the milled grinding matrix to beseparated from the particles comprising the biologically active materialcomprises particles of a size similar to and/or smaller than theparticles comprising the biologically active material, separationtechniques based on size distribution are inapplicable.

In these circumstances, the method of the present invention may involveseparation of at least a portion of the milled grinding matrix from thebiologically active material by techniques including but not limited toelectrostatic separation, magnetic separation, centrifugation (densityseparation), hydrodynamic separation, froth flotation.

Advantageously, the step of removing at least a portion of the milledgrinding matrix from the biologically active material may be performedthrough means such as selective dissolution, washing, or sublimation.

An advantageous aspect of the invention would be the use of grindingmatrix that has two or more components where at least one component iswater soluble and at least one component has low solubility in water. Inthis case washing can be used to remove the matrix component soluble inwater leaving the biologically active material encapsulated in theremaining matrix components. In a highly advantageous aspect of theinvention the matrix with low solubility is a functional excipient.

A highly advantageous aspect of the present invention is that certaingrinding matrixes appropriate for use in the method of the invention (inthat they physically degrade to the desired extent under dry millingconditions) are also pharmaceutically acceptable and thus appropriatefor use in a medicament. Where the method of the present invention doesnot involve complete separation of the grinding matrix from thebiologically active material, the present invention encompasses methodsfor the production of a medicament incorporating both the biologicallyactive material and at least a portion of the milled grinding matrix,medicaments so produced and methods of treatment of an animal, includingman, using a therapeutically effective amount of said biologicallyactive materials by way of said medicaments.

The medicament may include only the biologically active material and thegrinding matrix or, more preferably, the biologically active materialsand grinding matrix may be combined with one or more pharmaceuticallyacceptable carriers, as well as any desired excipients or other likeagents commonly used in the preparation of medicaments.

Analogously, a highly advantageous aspect of the present invention isthat certain grinding matrixes appropriate for use in the method of theinvention (in that they physically degrade to a desirable extent underdry milling conditions) are also appropriate for use in an agriculturalchemical composition. Where the method of the present invention does notinvolve complete separation of the grinding matrix from the biologicallyactive material, the present invention encompasses methods for theproduction of a agricultural chemical composition incorporating both thebiologically active material and at least a portion of the milledgrinding matrix, agricultural chemical composition so produced andmethods of use of such compositions.

The agricultural chemical composition may include only the biologicallyactive material and the grinding matrix or, more preferably, thebiologically active materials and grinding matrix may be combined withone or more acceptable carriers, as well as any desired excipients orother like agents commonly used in the preparation of agriculturalchemical compositions.

In one particular form of the invention, the grinding matrix is bothappropriate for use in a medicament and readily separable from thebiologically active material by methods not dependent on particle size.Such grinding matrixes are described in the following detaileddescription of the invention. Such grinding matrixes are highlyadvantageous in that they afford significant flexibility in the extentto which the grinding matrix may be incorporated with the biologicallyactive material into a medicament.

The mixture of biologically active material and grinding matrix may thenbe separated from the milling bodies and removed from the mill.

In one embodiment, the grinding matrix is separated from the mixture ofbiologically active material and grinding matrix. Where the grindingmatrix is not fully milled, the unmilled grinding matrix is separatedfrom the biologically active material. In a further aspect, at least aportion of the milled grinding matrix is separated from the biologicallyactive material.

The milling bodies are essentially resistant to fracture and erosion inthe dry milling process.

The quantity of the grinding matrix relative to the quantity ofbiologically active material, and the extent of milling of the grindingmatrix, is sufficient to provide reduced particle size of thebiologically active material.

The grinding matrix is neither chemically nor mechanically reactive withthe pharmaceutical material under the dry milling conditions of themethod of the invention except, for example, where the matrix isdeliberately chosen to undergo a mechanico-chemical reaction. Such areaction might be the conversion of a free base or acid to a salt orvice versa.

Preferably, the medicament is a solid dosage form, however, other dosageforms may be prepared by those of ordinary skill in the art.

In one form, after the step of separating said mixture of biologicallyactive material and grinding matrix from the plurality of millingbodies, and before the step of using said mixture of biologically activematerial and grinding matrix in the manufacture of a medicament, themethod may comprise the step of:

removing a portion of the grinding matrix from said mixture ofbiologically active material and grinding matrix to provide a mixtureenriched in the biologically active material;

and the step of using said mixture of biologically active material andgrinding matrix in the manufacture of a medicament, more particularlycomprises the step of using the mixture of biologically active materialand grinding matrix enriched in the biologically active material form inthe manufacture of a medicament.

The present invention includes medicaments manufactured by said methods,and methods for the treatment of an animal, including man, by theadministration of a therapeutically effective amount of the biologicallyactive materials by way of said medicaments.

In another embodiment of the invention, a facilitating agent or acombination of facilitating agents is also comprised in the mixture tobe milled. Such facilitating agents appropriate for use in the inventioninclude diluents, surfactants, polymers, binding agents, filling agents,lubricating agents, sweeteners, flavouring agents, preservatives,buffers, wetting agents, disintegrants, effervescent agents and agentsthat may form part of a medicament, including a solid dosage form, orother excipients required for other specific drug delivery, such as theagents and media listed below under the heading Medicinal andPharmaceutical Compositions, or any combination thereof.

Biologically Active Materials and Compositions

The present invention encompasses pharmaceutically acceptable materialsproduced according to the methods of the present invention, compositionsincluding such materials, including compositions comprising suchmaterials together with the grinding matrix with or without millingaids, facilitating agents, with at least a portion of the grindingmatrix or separated from the grinding matrix.

The pharmaceutically acceptable materials within the compositions of theinvention are present at a concentration of between about 0.1% and about99.0% by weight. Preferably, the concentration of pharmaceuticallyacceptable materials within the compositions will be about 5% to about80% by weight, while concentrations of 10% to about 50% by weight arehighly preferred. Desirably, the concentration will be in the range ofabout 10 to 15% by weight, 15 to 20% by weight, 20 to 25% by weight, 25to 30% by weight, 30 to 35% by weight, 35 to 40% by weight, 40 to 45% byweight, 45 to 50% by weight, 50 to 55% by weight, 55 to 60% by weight,60 to 65% by weight, 65 to 70% by weight, 70 to 75% by weight or 75 to80% by weight for the composition prior to any later removal (ifdesired) of any portion of the grinding matrix. Where part or all of thegrinding matrix has been removed, the relative concentration ofpharmaceutically acceptable materials in the composition may beconsiderably higher depending on the amount of the grinding matrix thatis removed. For example, if all of the grinding matrix is removed theconcentration of particles in the preparation may approach 100% byweight (subject to the presence of facilitating agents).

Compositions produced according to the present invention are not limitedto the inclusion of a single species of pharmaceutically acceptablematerials. More than one species of pharmaceutically acceptablematerials may therefore be present in the composition. Where more thanone species of pharmaceutically acceptable materials is present, thecomposition so formed may either be prepared in a dry milling step, orthe pharmaceutically acceptable materials may be prepared separately andthen combined to form a single composition.

Medicaments

The medicaments of the present invention may include thepharmaceutically acceptable material, optionally together with thegrinding matrix or at least a portion of the grinding matrix, with orwithout milling aids, facilitating agents, combined with one or morepharmaceutically acceptable carriers, as well as other agents commonlyused in the preparation of pharmaceutically acceptable compositions.

As used herein “pharmaceutically acceptable carrier” includes any andall solvents, dispersion media, coatings, antibacterial and antifungalagents, isotonic and absorption delaying agents, and the like that arephysiologically compatible. Preferably, the carrier is suitable forparenteral administration, intravenous, intraperitoneal, intramuscular,sublingual, pulmonary, transdermal or oral administration.Pharmaceutically acceptable carriers include sterile aqueous solutionsor dispersions and sterile powders for the extemporaneous preparation ofsterile injectable solutions or dispersion. The use of such media andagents for the manufacture of medicaments is well known in the art.Except insofar as any conventional media or agent is incompatible withthe pharmaceutically acceptable material, use thereof in the manufactureof a pharmaceutical composition according to the invention iscontemplated.

Pharmaceutical acceptable carriers according to the invention mayinclude one or more of the following examples:

-   -   (1) surfactants and polymers including, but not limited to        polyethylene glycol (PEG), polyvinylpyrrolidone (PVP),        polyvinylalcohol, crospovidone,        polyvinylpyrrolidone-polyvinylacrylate copolymer, cellulose        derivatives, hydroxypropylmethyl cellulose, hydroxypropyl        cellulose, carboxymethylethyl cellulose, hydroxypropylmethyl        cellulose phthalate, polyacrylates and polymethacrylates, urea,        sugars, polyols, and their polymers, emulsifiers, sugar gum,        starch, organic acids and their salts, vinyl pyrrolidone and        vinyl acetate    -   (2) binding agents such as various celluloses and cross-linked        polyvinylpyrrolidone, microcrystalline cellulose; and or    -   (3) filling agents such as lactose monohydrate, lactose        anhydrous, microcrystalline cellulose and various starches; and        or    -   (4) lubricating agents such as agents that act on the        flowability of the powder to be compressed, including colloidal        silicon dioxide, talc, stearic acid, magnesium stearate, calcium        stearate, silica gel; and or    -   (5) sweeteners such as any natural or artificial sweetener        including sucrose, xylitol, sodium saccharin, cyclamate,        aspartame, and acesulfame K; and or    -   (6) flavouring agents; and or    -   (7) preservatives such as potassium sorbate, methylparaben,        propylparaben, benzoic acid and its salts, other esters of        parahydroxybenzoic acid such as butylparaben, alcohols such as        ethyl or benzyl alcohol, phenolic chemicals such as phenol, or        quaternary compounds such as benzalkonium chloride; and or    -   (8) buffers; and or    -   (9) Diluents such as pharmaceutically acceptable inert fillers,        such as microcrystalline cellulose, lactose, dibasic calcium        phosphate, saccharides, and/or mixtures of any of the foregoing;        and or    -   (10) wetting agents such as corn starch, potato starch, maize        starch, and modified starches, croscarmellose sodium,        crospovidone, sodium starch glycolate, and mixtures thereof; and        or    -   (11) disintegrants; and or    -   (12) effervescent agents such as effervescent couples such as an        organic acid (e.g., citric, tartaric, malic, fumaric, adipic,        succinic, and alginic acids and anhydrides and acid salts), or a        carbonate (e.g. sodium carbonate, potassium carbonate, magnesium        carbonate, sodium glycine carbonate, L-lysine carbonate, and        arginine carbonate) or bicarbonate (e.g. sodium bicarbonate or        potassium bicarbonate); and or    -   (13) other pharmaceutically acceptable excipients.

Medicaments of the invention suitable for use in animals and inparticular in man typically must be stable under the conditions ofmanufacture and storage. The medicaments of the invention comprising thebiologically active material can be formulated as a solid, a solution, amicroemulsion, a liposome, or other ordered structures suitable to highdrug concentration. Actual dosage levels of the biologically activematerial in the medicament of the invention may be varied in accordancewith the nature of the biologically active material, as well as thepotential increased efficacy due to the advantages of providing andadministering the biologically active material (e.g., increasedsolubility, more rapid dissolution, increased surface area of thebiologically active material, etc.). Thus as used herein“therapeutically effective amount” will refer to an amount ofbiologically active material required to effect a therapeutic responsein an animal. Amounts effective for such a use will depend on: thedesired therapeutic effect; the route of administration; the potency ofthe biologically active material; the desired duration of treatment; thestage and severity of the disease being treated; the weight and generalstate of health of the patient; and the judgment of the prescribingphysician.

In another embodiment, the biologically active material, optionallytogether with the grinding matrix or at least a portion of the grindingmatrix, of the invention may be combined into a medicament with anotherbiologically active material, or even the same biologically activematerial. In the latter embodiment, a medicament may be achieved whichprovides for different release characteristics—early release from thebiologically active material, and later release from a larger averagesize biologically active material.

Pharmacokinetic Properties of Diclofenac Compositions

Suitable animal models to determine pharmacokinetic parameters aredescribed in the prior art, such as the beagle dog model described inU.S. Pat. No. 7,101,576.

Fast Onset of Activity

The diclofenac compositions of the invention exhibit faster therapeuticeffects.

In one example, following administration the diclofenac compositions ofthe invention comprising diclofenac have a T_(max) of less than about 5hours, less than about 4.5 hours, less than about 4 hours, less thanabout 3.5 hours, less than about 3 hours, less than about 2.75 hours,less than about 2.5 hours, less than about 2.25 hours, less than about 2hours, less than about 1.75 hours, less than about 1.5 hours, less thanabout 1.25 hours, less than about 1.0 hours, less than about 50 minutes,less than about 40 minutes, less than about 30 minutes, less than about25 minutes, less than about 20 minutes, less than about 15 minutes, lessthan about 10 minutes, less than about 5 minutes, or less than about 1minute.

Increased Bioavailability

The diclofenac compositions of the invention preferably exhibitincreased bioavailability (AUC) and require smaller doses as compared toprior conventional compositions administered at the same dose. Any drugcomposition can have adverse side effects. Thus, lower doses of drugswhich can achieve the same or better therapeutic effects as thoseobserved with larger doses of conventional compositions are desired.Such lower doses can be realized with the compositions of the inventionbecause the greater bioavailability observed with the compositions ascompared to conventional drug formulations means that smaller doses ofdrug are required to obtain the desired therapeutic effect.

The Pharmacokinetic Profiles of the Compositions of the Invention arenot Substantially Affected by the Fed or Fasted State of the SubjectIngesting the Compositions

The invention encompasses diclofenac compositions wherein thepharmacokinetic profile of the composition is not substantially affectedby the fed or fasted state of a subject ingesting the composition. Thismeans that there is no substantial difference in the quantity ofcomposition or the rate of composition absorption when the compositionsare administered in the fed versus the fasted state. Thus, thecompositions of the invention substantially eliminate the effect of foodon the pharmacokinetics of the composition.

The difference in absorption of the diclofenac composition of theinvention, when administered in the fed versus the fasted state, is lessthan about 35%, less than about 30%, less than about 25%, less thanabout 20%, less than about 15%, less than about 10%, less than about 5%,or less than about 3%. This is an especially important feature intreating patients with difficulty in maintaining a fed state.

In addition, preferably the difference in the rate of absorption (i.e.,T_(max)) of the diclofenac compositions of the invention, whenadministered in the fed versus the fasted state, is less than about100%, less than about 90%, less than about 80%, less than about 70%,less than about 60%, less than about 50%, less than about 40%, less thanabout 30%, less than about 20%, less than about 15%, less than about10%, less than about 5%, less than about 3%, or essentially nodifference. Benefits of a dosage form which substantially eliminates theeffect of food include an increase in subject convenience, therebyincreasing subject compliance, as the subject does not need to ensurethat they are taking a dose either with or without food.

Preferably, the T_(max) of an administered dose of a diclofenaccomposition of the invention is less than that of a conventional drugactive composition, administered at the same dosage.

A preferred diclofenac composition of the invention exhibits incomparative pharmacokinetic testing with a standard conventional drugactive composition, in oral suspension, capsule or tablet form, aT_(max) which is less than about 100%, less than about 90%, less thanabout 80%, less than about 70%, less than about 60%, less than about50%, less than about 40%, less than about 30%, less than about 25%, lessthan about 20%, less than about 15%, or less than about 10% of theT_(max) exhibited by the standard conventional drug active composition.

In addition, preferably the C_(max) of a diclofenac composition of theinvention is greater than the C_(max) of a conventional drug activecomposition, administered at the same dosage. A preferred diclofenaccomposition of the invention exhibits in comparative pharmacokinetictesting with a standard conventional drug active composition, in oralsuspension, capsule or tablet form, a C_(max) which is greater thanabout 5%, greater than about 10%, greater than about 15%, greater thanabout 20%, greater than about 30%, greater than about 40%, greater thanabout 50%, greater than about 60%, greater than about 70%, greater thanabout 80%, greater than about 90%, greater than about 100%, greater thanabout 110%, greater than about 120%, greater than about 130%, greaterthan about 140%, or greater than about 150% than the C_(max) exhibitedby the standard conventional drug active composition.

In addition, preferably the diclofenac composition has an AUC greaterthan that of the equivalent conventional composition administered at thesame dosage. A preferred diclofenac composition of the inventionexhibits in comparative pharmacokinetic testing with a standardconventional drug active composition, in oral suspension, capsule ortablet form, a AUC which is greater than about 5%, greater than about10%, greater than about 15%, greater than about 20%, greater than about30%, greater than about 40%, greater than about 50%, greater than about60%, greater than about 70%, greater than about 80%, greater than about90%, greater than about 100%, greater than about 110%, greater thanabout 120%, greater than about 130%, greater than about 140%, or greaterthan about 150% than the AUC exhibited by the standard conventional drugactive composition.

Any standard pharmacokinetic protocol can be used to determine bloodplasma concentration profile in humans following administration of acomposition, and thereby establish whether that composition meets thepharmacokinetic criteria set out herein. For example, a randomizedsingle-dose crossover study can be performed using a group of healthyadult human subjects. The number of subjects should be sufficient toprovide adequate control of variation in a statistical analysis, and istypically about 10 or greater, although for certain purposes a smallergroup can suffice. Each subject receives by oral administration at timezero a single dose (e.g., 300 mg) of a test formulation of composition,normally at around 8 am following an overnight fast. The subjectscontinue to fast and remain in an upright position for about 4 hoursafter administration of the composition. Blood samples are collectedfrom each subject prior to administration (e.g., 15 minutes) and atseveral intervals after administration. For the present purpose it ispreferred to take several samples within the first hour, and to sampleless frequently thereafter. Illustratively, blood samples could becollected at 15, 30, 45, 60, and 90 minutes after administration, thenevery hour from 2 to 10 hours after administration. Additional bloodsamples may also be taken later, for example at 12 and 24 hours afteradministration. If the same subjects are to be used for study of asecond test formulation, a period of at least 7 days should elapsebefore administration of the second formulation. Plasma is separatedfrom the blood samples by centrifugation and the separated plasma isanalyzed for composition by a validated high performance liquidchromatography (HPLC) or liquid chromatography mass spectrometry (LCMS)procedure. Plasma concentrations of composition referenced herein areintended to mean total concentrations including both free and boundcomposition.

Any formulation giving the desired pharmacokinetic profile is suitablefor administration according to the present methods. Exemplary types offormulations giving such profiles are liquid dispersions and solid doseforms of composition. If the liquid dispersion medium is one in whichthe composition has very low solubility, the particles are present assuspended particles. The smaller the particles the higher theprobability that the formulation will exhibit the desiredpharmacokinetic profile.

Thus, an diclofenac composition of the invention, upon administration toa subject, provides improved pharmacokinetic and/or pharmacodynamicproperties compared with a standard reference diclofenac composition asmeasured by at least one of speed of absorption, dosage potency,efficacy, and safety.

Modes of Administration of Medicaments Comprising Biologically ActiveMaterials

Medicaments of the invention can be administered to animals, includingman, in any pharmaceutically acceptable manner, such as orally,rectally, pulmonary, intravaginally, locally (powders, ointments ordrops), transdermal, parenteral administration, intravenous,intraperitoneal, intramuscular, sublingual or as a buccal or nasal spray

Solid dosage forms for oral administration include capsules, tablets,pills, powders, pellets, and granules. Further, incorporating any of thenormally employed excipients, such as those previously listed, andgenerally 5-95% of the biologically active agent, and more preferably ata concentration of 10%-75% will form a pharmaceutically acceptablenon-toxic oral composition.

Medicaments of the invention may be parenterally administered as asolution of the biologically active agent suspended in an acceptablecarrier, preferably an aqueous carrier. A variety of aqueous carriersmay be used, e.g. water, buffered water, 0.4% saline, 0.3% glycine,hyaluronic acid and the like. These compositions may be sterilized byconventional, well known sterilization techniques, or may be sterilefiltered. The resulting aqueous solutions may be packaged for use as is,or lyophilized, the lyophilized preparation being combined with asterile solution prior to administration.

For aerosol administration, medicaments of the invention are preferablysupplied along with a surfactant or polymer and propellant. Thesurfactant or polymer must, of course, be non-toxic, and preferablysoluble in the propellant. Representative of such agents are the estersor partial esters of fatty acids containing from 6 to 22 carbon atoms,such as caproic, octanoic, lauric, palmitic, stearic, linoleic,linolenic, olesteric and oleic acids with an aliphatic polyhydricalcohol or its cyclic anhydride. Mixed esters, such as mixed or naturalglycerides may be employed. The surfactant or polymer may constitute0.1%-20% by weight of the composition, preferably 0.25-5%. The balanceof the composition is ordinarily propellant. A carrier can also beincluded, as desired, as with, e.g., lecithin for intranasal delivery.

Medicaments of the invention may also be administered via liposomes,which serve to target the active agent to a particular tissue, such aslymphoid tissue, or targeted selectively to cells. Liposomes includeemulsions, foams, micelles, insoluble monolayers, liquid crystals,phospholipid dispersions, lamellar layers and the like. In thesepreparations the composite microstructure composition is incorporated aspart of a liposome, alone or in conjunction with a molecule that bindsto or with other therapeutic or immunogenic compositions.

As described above, the biologically active material can be formulatedinto a solid dosage form (e.g., for oral or suppository administration),together with the grinding matrix or at least a portion of it. In thiscase there may be little or no need to add stabilizing agents since thegrinding matrix may effectively act as a solid-state stabilizer.

However, if the biologically active material is to be utilized in aliquid suspension, the particles comprising the biologically activematerial may require further stabilization once the solid carrier hasbeen substantially removed to ensure the elimination, or at leastminimisation of particle agglomeration.

Therapeutic Uses

Therapeutic uses of the medicaments of the invention include painrelief, anti-inflammatory, migraine, asthma, and other disorders thatrequire the active agent to be administered with a high bioavailability.

One of the main areas when rapid bioavailability of a biologicallyactive material is required is in the relief of pain. The minoranalgesics, such as cyclooxgenase inhibitors (aspirin related drugs) maybe prepared as medicaments according to the present invention.

Medicaments of the invention may also be used for treatment of eyedisorders. That is, the biologically active material may be formulatedfor administration on the eye as an aqueous suspension in physiologicalsaline, or a gel. In addition, the biologically active material may beprepared in a powder form for administration via the nose for rapidcentral nervous system penetration.

Treatment of cardiovascular disease may also benefit from biologicallyactive materials according to the invention, such as treatment of anginapectoris and, in particular, molsidomine may benefit from betterbioavailability.

Other therapeutic uses of the medicaments of the present inventioninclude treatment of hair loss, sexual dysfunction, or dermal treatmentof psoriasis.

The present invention will now be described with reference to thefollowing non-limiting Examples. The description of the Examples is inno way limiting on the preceding paragraphs of this specification, butis provided for exemplification of the methods and compositions of theinvention.

EXAMPLES

It will be apparent to persons skilled in the milling and pharmaceuticalarts that numerous enhancements and modifications can be made to theabove described processes without departing from the basic inventiveconcepts. For example, in some applications the biologically activematerial may be pretreated and supplied to the process in the pretreatedform. All such modifications and enhancements are considered to bewithin the scope of the present invention, the nature of which is to bedetermined from the foregoing description and the appended claims.Furthermore, the following Examples are provided for illustrativepurposes only, and are not intended to limit the scope of the processesor compositions of the invention.

The Following Materials were Used in the Examples

Active pharmaceutical ingredients were sourced from commercialsuppliers, excipients from either commercial suppliers such asSigma-Aldrich or retailers, while food ingredients were sourced fromretailers.

The Following Mills were Used for the Grinding Experiments

Spex-Type Mill:

Small scale milling experiments were conducted using a vibratory Spex8000D mixer/mill. Twelve ⅜″ stainless steel balls were used as thegrinding media. The powder charge and grinding media were loaded into ahardened steel vial with an internal volume of approximately 75 mL.Following milling, the milled material was discharged from the vial andsieved to remove grinding media.

Attritor-Type Mill:

Small scale attritor milling experiments were performed using a 1HDUnion Process attritor mill with a 110 mL grinding chamber. The grindingmedia consisted of 330 g 5/16″ stainless steel balls. The mill wasloaded through the loading port, with dry materials added initially,followed by the grinding media. The milling process was conducted withthe jacket cooled at 10-20° C. and the shaft rotating at 500 rpm. Uponcompletion of milling, the milled material was discharged from the milland sieved to remove the grinding media.

Medium scale attritor milling experiments were performed using a 1HDUnion Process attritor mill with a 1 L grinding chamber or a 1S UnionProcess attritor mill with a 750 mL grinding chamber. The grinding mediaconsisted of 3 kg of 5/16″ stainless steel balls or 1.5 kg of ⅜″stainless steel balls for the 1S attritor. The 1HD mill was loadedthrough the loading port, with dry materials added initially, followedby the grinding media, while the grinding media was added initially,followed by the dry materials in the 1S attritor mill. The millingprocess was conducted with the jacket cooled at 10-20° C. with the shaftrotating at 350 rpm in the 1HD attritor or 550 rpm in the 1S attritor.Upon completion of milling, the milled material was discharged from themill and sieved to remove the grinding media.

Medium to large scale attritor milling experiments were performed usinga 1S Union Process attritor mill with a ½ gallon grinding chamber. Thegrinding media consisted of 7 kg of ⅜″ stainless steel balls. The millwas loaded through the loading port, with the grinding media addedinitially, followed by the dry powders. The milling process wasconducted with the jacket cooled at 18° C. and the shaft rotating at550-555 rpm. Upon completion of milling, the milled powder wasdischarged from the mill through the bottom discharge port at 77 rpm for5 min.

Large scale attritor milling experiments were performed using a 1S UnionProcess attritor mill with a 1½ gallon grinding chamber. The grindingmedia consisted of 20 kg of ⅜″ stainless steel balls. The mill wasloaded through the loading port, with the grinding media addedinitially, then followed by the dry powders. The milling process wasconducted with the jacket cooled to ambient temperature and the shaftrotating at 300 rpm. Upon completion of milling, the milled powder wasdischarged from the mill through the bottom discharge port at 77 rpm for5 min. The largest scale attritor millings were done in a 30S UnionProcess mill with a 25 gallon grinding chamber (Union Process, AkronOhio, USA). The grinding media consisted of 454 kg of ⅜″ stainless steelballs. The mill was loaded through its split top lid, with the grindingmedia added initially, then followed by the dry powders (25 kg). Themilling process was conducted with the jacket cooled to 10° C. and theshaft rotating at 130 rpm. Upon completion of milling, the milled powderwas discharged from the mill through the bottom discharge port at 77 rpmfor 5 min.

Siebtechnik Mill

Medium scale milling experiments were also performed in a SiebtechnikGSM06 (Siebtechnik, GmbH, Germany) with two 1 L milling chambers. Eachchamber was filled with 2.7 kg stainless steel media with a diameter of⅜″. The media and powder were loaded with the lid off. The mill wasoperated at ambient temperature. The vibration speed was the standardmill settings. Upon completion of the milling the media was separatedfrom the powder by sieving.

Simoloyer Mill

Medium scale milling experiments were performed in a Simoloyer CM01 (ZOZGmbH, Germany) with a 2 L milling chamber. The grinding media consistedof 2.5 kg stainless steel media with a diameter of 5 mm. the media wasloaded though the loading port followed by the dry materials. Themilling vessel was cooled using water at a temperature of about 18° C.The mill speed was operated in cycle mode: at 1300 rpm for two minutesand at 500 rpm for 0.5 min and so forth. Upon completion of the millingthe media was discharged from the mill using a grated valve to retainthe grinding media.

Large scale milling experiments were performed in a Simoloyer CM100 (ZOZGmbH, Germany) with a 100 L milling chamber. The grinding mediaconsisted of 100 kg stainless steel media with a diameter of 3/16″. Thepowder charge (11 kg) was added to the milling chamber, which alreadycontained the grinding media, through a loading port. The millingchamber was cooled to 18° C. and the powder was milled for a total of 20minutes using a cycling mode equivalent to a tip speed at 1300/500 rpmfor 2/0.5 min in the CM-01 type mill. Upon completion of the milling themill was discharged by sucking the powder into a cyclone.

Hicom Mill

Millings performed in a nutating Hicom mill utilized 14 kg of stainlesssteel 0.25″ grinding media together with a powder charge of 480 g. Themill was loaded by pre-mixing media and powder, then adding the mixtureto the grinding chamber through the loading port at the top of the mill.The milling was done at 1000 rpm and the mill discharged by invertingthe mill and emptying through the loading port. The recovered materialwas sieved to separate the grinding media from the powder.

Variations to the milling conditions set out above are indicated in thevariations column in the data tables. The key to these variations isshown in Table A.

Particle Size Measurement:

The particle size distribution (PSD) was determined using a MalvernMastersizer 2000 fitted with a Malvern Hydro 2000S pump unit.Measurement settings used: Measurement Time: 12 seconds, Measurementcycles: 3. Final result generated by averaging the 3 measurements.Samples were prepared by adding 200 mg of milled material to 5.0 mL of1% PVP in 10 mM hydrochloric acid (HCl), vortexing for 1 min and thensonicating. From this suspension enough was added into the dispersant(10 mM HCl) to attain a desired obscuration level. If necessary an extra1-2 minutes of sonication was applied using the internal sonicationprobe in the measurement cell. The refractive index of the activeingredient to be measured was in the range of 1.49-1.73. Any variationsto this general method are summarized in Table B.

XRD Analysis:

Powder X-Ray diffraction (XRD) patterns were measured with aDiffractometer D 5000, Kristalloflex (Siemens). The measurement rangewas from 5-18 degrees 2-Theta. The slit width was set to 2 mm and thecathode ray tube was operated at 40 kV and 35 mA. Measurements wererecorded at room temperature. The recorded traces were subsequentlyprocessed using Bruker EVA software to obtain the diffraction pattern.

TABLE A Variations to milling conditions. Only conditions reported inthe table have changed as compared to conditions reported above. MillMilling Media Media Offload Variation # type Speed (rpm) size (inch)Mass (kg) spped (rpm) A 1HD 1L 0.25 B 1S 0.5 gal 5 C 1S 0.5 gal 4 D 1S0.5 gal 500 E 1S 0.5 gal 550-555 F 1S 1.5 gal 316-318 21 G 1S 1.5 gal500 21 H 1S 1.5 gal 355 21 I 1S 1.5 gal 355 18 J 1S 1.5 gal 21 K 1S 1.5gal 18.4 L 1S 1.5 gal 400 M 1S 1.5 gal 21 57 N 1S 1.5 gal 57 0 1S 0.5gal 400 400 P 1S 0.5 gal 500 350 Q HICOM 1/8 R HICOM 11.7

TABLE B Variations to particle size measurement conditions. SampleMeasurement Addition Variation # Dispersant Dispersant Method 1 0.1%PVPin DI water Powder addition 2 0.2% Pluronic L81 DI water in DI water 3Saturated glyphosate Powder in DI water addition 4 Saturated glyphosatePowder in DI water addition 5 1% PVP in DI water DI water 6 DI waterPowder addition 7 1% PVP in DI water Saturated creatine in DI water 8 1%PVP in DI water 10 mM HCl 9 0.2% Pluronic L81 Acidified with 1M HCl inDI water 10 1% PVP in DI water 0.1% PVP in DI water 11 1% PVP in DIwater 1% PVP in DI water 12 Filtered before PSD measurement

ABBREVIATIONS

HCl: Hydrochloric acid

Nap: Naproxen acid

PSD: Particles size distribution

PVP: Polyvinyl pyrrolidone

RI: Refractive index

Rpm: Revolutions per minute

SLS: Sodium lauryl sulphate

SSB: Stainless Steel Balls

XRD: X-Ray Diffraction

Other abbreviations used in the data tables are listed below in Table C(for actives), Table D (for matrices) and Table E (for surfactants). Inthe data tables single letter with example number abbreviations havebeen used to identify specific sample numbers within the table. The datatables shown in the figures the use of surfactant, matrix areinterchangeable and do not necessarily define the nature of thatmaterial.

TABLE C Abbreviations used for active pharmaceutical ingredients. APIName Abbreviation 2,4-Dichlorophenoxyacetic 2,4D acid Anthraquinone ANTCelecoxib CEL Cilostazol CIL Ciprofloxacin CIP Creatine Monohydrate CRMCyclosporin A CYA Diclofenac Acid DIC Glyphosate GLY Halusulfuron HALDiclofenac IND Mancozeb MAN Meloxicam MEL Metaxalone MTX Metsulfuron METNaproxen Acid NAA Naproxen Sodium NAS Progesterone PRO Salbutamol SALSulfur SUL Tribenuran TRI

TABLE D Abbreviations used for excipients. Matrix Name AbbreviationCalcium Carbonate CAC Glucose GLU Lactose Anhydrous LAA LactoseMonohydrate LAC Lactose Monohydrate Food LFG Grade Malic Acid MAAMaltitol MAL Mannitol MAN Sodium Bicarbonate SB Sodium Chloride SCSorbitol SOR Sucrose SUC Tartaric Acid TA TriSodium Citrate DihydrateTCD Whey Powder WP Xylitol XYL

TABLE E Abbreviations used for surfactants Surfactant Name AbbreviationAerosil R972 Silica AS Benzalkonium Chloride BC Brij700 B700 Brij76 B76Cremophor EL CEL Cremophor RH-40 C40 Dehscofix 920 D920 Docusate SodiumDS Kollidon 25 K25 Kraftsperse 1251 K1251 Lecithin LEC Poloxamer 188P188 Microcrystalline Cellulose MCC Poloxamer 407 P407 PolyethyleneGlycol 3000 P3000 Polyethylene Glycol 8000 P8000 Polyoxyethylene 40Stearate P4OS Polyvinyl Pyrrolidone (Kollidon 30) PVP Primellose PMLPrimojel PRI Sodium Deoxycholate SDC Sodium Dodecyl Sulphate SDS SodiumDodecylbenzenesulphonic SDA Acid Sodium N-Lauroyl Sarcosine SNS SodiumOctadecyl Sulphate SOS Sodium Pentane Sulphonate SPS Soluplus HS15 SOLTeric 305 T305 Tersperse 2700 T2700 Terwet 1221 T1221 Terwet 3785 T3785Tween 80 T80

Example 1 Spex Milling

A range of actives, matrices and surfactants in a variety ofcombinations were milled using the Spex mill. The details of thesemillings are shown in FIGS. 1A-1G together with the particle sizedistributions of actives that were milled.

These millings demonstrate that the addition of a small amount ofsurfactant to the milling matrix delivers a smaller particle sizecompared to millings of just an active and a single matrix. Someexamples of this are samples Z and AA compared to sample Y; Sample ABcompared to sample AC; sample AE compared to sample AD; sample AGcompared to sample AF; sample AP compared to sample AO; sample ARcompared to sample AQ, sample AT compared to sample AS; Samples AX, AYand AZ compared to sample AW; sample BC compared to sample BD; sample BIcompared to BH; samples BL-BR compared to sample BK; samples CS-DBcompared to sample DC. This last example is particularly noteworthy asthese millings were undertaken at 45% v/v. This demonstrates the broadapplicability of this invention. Some other examples of surfactantaddition being beneficial for size reduction are samples DD-DG and DI-DKcompared to sample DH; sample DM compared to sample DL. Other samplessuch as samples DY-EC compared to sample DX; sample AV compared tosample AU; samples B-H compared to sample A and samples K-M compared tosample J show this ti be also true when particle size statistics suchthe %<1 micron as used.

Note that this applies to mechanochemical matrix milling as well. Thisis demonstrated by sample BI where naproxen sodium is milled withtartaric acid and converted to naproxen acid. FIG. 1H shows XRD datathat demonstrates the transformation.

Other samples such as CB-CR show examples were surfactants suitable foruse with IV formulations can be used to manufacture very smallparticles.

It is also noteworthy that samples DS and DT could be sized using asaturated solution of the active (salbutamol) demonstrating that activeswith high water solubility can be measured as long as care is taken whenmeasuring the size.

Two sets of data, samples N-Q and samples R-U, also demonstrate that theinvention described herein is unique. In these samples the active milledwith a matrix and surfactant produces small particles. When milled withmatrix alone the particles sizes are larger, in the case of sample Qthey are not even nanoparticles. When the active is milled with just 1%surfactant the resultant particle size is very large. Even when 80%surfactant is used the size is large.

Example 2 110 mL Attritor

A range of actives, matrices and surfactants in a variety ofcombinations were milled using the 110 ml stirred attritor mill. Thedetails of these millings are shown in FIG. 2A together with theparticle size distributions of actives that were milled.

These millings also demonstrate that the addition of a small amount ofsurfactant to the milling matrix delivers a smaller particle sizecompared to millings of just an active and a single matrix in a smallscale stirred mill as well as the vibratory Spex mill. Sample F alsodemonstrates that small particles can be achieved at high % actives whena surfactant is present. Sample D and E also show that the addition ofthe surfactant also increased the yield of powder from the mill.

Example 3 Second Matrix

In this example naproxen was milled with a mixture of two matrices usingthe Spex mill. The details of these millings are shown in FIG. 3Atogether with the particle size distributions of actives that weremilled. Samples A and B were milled in a primary matrix of lactosemonohydrate and 20% of second matrix. The particle size of thesemillings is smaller than the same milling with just lactose monohydrate(See example 1 sample No AH, FIG. 1B). The particle size is also smallerthan naproxen milled in the secondary matrices (See example 1 sample NoAI and AJ, FIG. 1B). This shows the mixed matrices have synergytogether.

Samples C-E were milled in anhydrous lactose with 20% of a secondmatrix. All these samples had a particle size much smaller than naproxenmilled in anhydrous lactose alone (See example 1 sample No AK, FIG. 1B).

These millings demonstrate that the addition of a second matrix to theprimary milling matrix delivers a smaller particle size compared tomillings with just a single matrix.

Example 4 1 L Attritor

Two actives with various combinations of lactose monohydrate and SDSwere milled using the 1 L stirred attritor mill. The details of thesemillings are shown in FIG. 4A together with the particle sizedistributions of actives that were milled.

Sample A and B are millings of meloxicam at 20%. While sample B has aslightly smaller particle size than sample A there is a dramaticdifference in the amount of material recovered from the milling. SampleA, milled with 3% SDS has a high yield of 90% whereas sample B with nosurfactant has practically no yield with all the powder caked in themill.

In samples C-F the milling of 13% diclofenac shows that the use of asecond matrix (tartaric acid) in combination with 1% SDS delivers thebest outcome of a good particle size and high yield. Sample D which hasjust the mixed matrix has very good particle size but poor yield.

These results show that the addition of a small amount of surfactantimproves milling performance.

Example 5 750 mL Attritor

Two actives with various combinations surfactants were milled using the750 ml stirred attritor mill. The details of these millings are shown inFIG. 5A together with the particle size distributions of actives thatwere milled.

In samples A-C three millings of naproxen are shown. Sample A has just1% SDS as a surfactant. Samples B and C have a second surfactant presentand these samples have a smaller particle size as measured by the %<500nm, %<1000 nm and %<2000 nm.

In samples D-F three millings of diclofenac are shown. Sample D has just1% SDS as a surfactant. Samples E and F have a second surfactant presentand these samples have a smaller particle size compared to sample D.

These examples demonstrate that the use of combination of surfactantscan be useful to achieve better reduction in particle size.

Example 6 ½ Gallon 1S

A range of actives, matrices and surfactants in a variety ofcombinations were milled using the ½ gallon 1S mill. The details ofthese millings are shown in FIGS. 6A-C together with the particle sizedistributions of actives that were milled.

The following examples demonstrate the increased yield obtained whenmilling an active in a ½ gallon 1S attritor mill with a surfactant ascompared to no surfactant, with all other factors being identical.Sample C and D (FIG. 6A) shows Naproxen acid milled in Mannitol withyields of 92% and 23%, with and without surfactant. Sample S and AL(FIGS. 6B and C) show the same for glyphosate with yields of 95% and26%, respectively. Sample AI and AJ (FIG. 6B) show Ciprofloxacin yieldsof 94% and 37% with and without surfactant while sample AM an AN (FIG.6C) show Celecoxib yields of 86% and 57% with and without surfactants.Finally, samples AP and AQ (FIG. 6C) shows milling Mancozeb with orwithout surfactants results in yields of 90% and 56%, respectively.

The following examples illustrates that milling an active in a ½ gallon1S attritor mill with a surfactant as compared to without surfactant andall other factors identical, leads to smaller particle size aftermilling. Sample C and D (FIG. 6A) shows a D(0.5) of 0.181 and 0.319 withor without surfactant, while sample AM and AN (FIG. 6C) shows D(0.5) of0.205 and 4.775 with and without surfactants.

The series of samples Q-S are timepoints taken from a single glyphosatemilling. The data demonstrates that the size of the actives decreaseswith milling time.

Other samples such as V-AA show examples were surfactants suitable foruse with IV formulations can be used to manufacture very smallparticles.

Some of the particle size data in FIGS. 6A-C was converted to a numberaverage particle size and is shown in the tables. This number wascalculated in the following way. The Volume distribution was transformedto the number distribution using the Malvern Mastersizer software. Foreach size bin the size of the bin was multiplied by the % of particlesin the bin. This numbers were added together and divided by 100 to givethe number average particle size.

Example 7 Metaxalone

Metaxalone was milled with various combinations of matrices andsurfactants using a variety of mills. The details of these millings areshown in FIG. 7A together with the particle size distributions ofactives that were milled. Samples A, B, E, G, H and I were milled in aSpex mill. Samples C, D and F were milled in the 750 ml attritor. Theremaining samples were milled in the ½ gallon 1S mill.

Samples A compared to sample B and sample H compared to sample Gdemonstrate that the addition of one or more surfactants enables theproduction of smaller active particles. Other millings such as samplesC-F show that metaxalone can be milled small at very high activeloadings. Sample I shows that disintegrant can be added during millingand not effect the production of small active particles. Note that theparticle size in sample I is after filtration through a 10 micronfilter. Sample N shows an alternative way to manufacture a formulationwith small particles and disintegrants. In this example the powder fromsample M was left in the mill and a wetting agent (PVP) and disintegrantwere added. The powder was milled for a further 2 minutes and thenunloaded with a very high yield of 97%.

The series of samples J-M are timepoints taken from a single milling.The data demonstrates that the size of the actives decreases withmilling time.

Example 8 Hicom

A range of actives, matrices and surfactants in a variety ofcombinations were milled using the Hicom mill. The details of thesemillings are shown in FIG. 8A together with the particle sizedistributions of actives that were milled.

The data shows that the invention described herein can be used with theHicom mill with its nutating action. The data in FIG. 8A shows that avariety of actives can be milled small in very short times and give verygood yields at 500 gram scale.

Sample N and 0 show that cocoa powder can be reduced to very fine sizesin short times using the invention describes here in combination withthe Hicom nutating mill. Likewise Sample P shows that this is also thecase for cocoa nibs.

Example 9 1.5 Gallon 1S

A range of actives, matrices and surfactants in a variety ofcombinations were milled using the 1.5 Gallon 1S mill. The details ofthese millings are shown in FIGS. 9A-B together with the particle sizedistributions of actives that were milled.

The following examples demonstrate the increased yield obtained whenmilling an active in a 1.5 gallon 1S attritor mill with a surfactant ascompared to no surfactant, with all other factors being identical.Sample J and N (FIG. 9A) shows yields of 51% and 80%, without and withsurfactant. Sample K and P (FIG. 9A) show yields of 27% and 80%, withoutand with surfactant, while sample L (FIG. 9A) show a yield of 94% withsurfactant and the control without surfactant (sample M, FIG. 9A)resulted in no yield due to caking within the mill.

The following examples illustrates that milling an active in a 1.5gallon 1S attritor mill with a surfactant as compared to withoutsurfactant and all other factors identical, leads to smaller particlesize after milling. Sample F and G (FIG. 9A) shows a D(0.5) of 0.137 and4.94 with or without surfactant, while sample K and P (FIG. 9A) showsD(0.5) of 0.242 and 0.152 without and with surfactants.

The series of samples AI-AL are timepoints taken from a single meloxicammilling. The data demonstrates that the size of the actives decreaseswith milling time.

Other samples such as A-E show examples were surfactants suitable foruse with IV formulations can be used to manufacture very smallparticles.

Sample M was a milling of meloxicam in lactose monohydrate withoutsurfactant. 3 minutes into the milling the mill refused to turn. Themilling was stopped and started again but only ran for another 3 minutesbefore stopping again. At this point the mill was taken apart and noevidence of caking was found. However the powder had a gritty feeling toit and was locking the medium and shaft such that it was not possible toturn. The media was weighed and it as found that 150 grams of powder wason the media indicating that it was sticking to the media and making ithard to move. At this point the mill was re-assembled and the powder andmedia put back in. 30.4 grams of SDS was included in the milling makingit similar to milling L. After the addition of the surfactant the millwas run for another 14 minutes (giving a total of 20 mins) withoutincident. After offloading the powder the media was weighed and theweigh of powder on the media was only 40.5 grams. This indicates theaddition of surfactant has improved the milling performance and abilityto mill the powder.

Some of the particle size data in FIGS. 9A-B was converted to a numberaverage particle size and is shown in the tables. This number wascalculated in the following way. The Volume distribution was transformedto the number distribution using the Malvern Mastersizer software. Foreach size bin the size of the bin was multiplied by the % of particlesin the bin. This numbers were added together and divided by 100 to givethe number average particle size.

Example 10 Large scale 25/11 kg

Sample A (FIG. 10A) was milled in the Siebtechnik mill for 15 minutes.After this time the powder was completely caked onto the walls of themill and the media. No powder could be removed to measure the particlesize. At this point 0.25 g (1 w/w %) SLS was added to mill chamber andmilling was then undertaken for a further 15 minutes. After the secondperiod of milling in the presence of SLS powder was no longer caked ontothe media and some free powder was also present. The observations madebefore and after the addition of the SLS demonstrate that the additionof the surfactant lessens the problem of caking. With the addition ofsurfactant the caked material could be recovered to become free powderagain with small particle size.

Sample B-E was milled in horizontal Simoloyer mills. The details ofthese millings are shown in FIG. 10A together with the particle sizedistributions of actives that were milled.

The data shows that the invention described herein can be used withSimoloyer mills with their horizontal attritor action. Of particularnote is example E which was milled at 11 kg scale. This demonstrates theinvention described herein is suitable for commercial scale milling.

Sample F was milled in a vertical attritor mill (Union Process S-30).The details of this milling is shown in FIG. 10A together with theparticle size distribution of the active milled.

The data shows that the invention described herein can be used with aS-30 mills with its vertical attritor action. Of particular note is thatthis milling was at 25 kg scale. This demonstrates the inventiondescribed herein is suitable for commercial scale milling.

Example 11 Naproxen

Naproxen was milled in mannitol with a range of surfactants using the ½Gallon 1S mill. The details of these millings are shown in FIG. 11Atogether with the particle size distributions of actives that weremilled.

Naproxen acid milled in Mannitol with a surfactant (Sample A, D-J inFIG. 11A) leads to higher yields, as compared to Naproxen acid milled inMannitol without surfactant (Sample K, FIG. 11A). Naproxen acid milledin Mannitol and either microcrystalline cellulose or the disintegrantprimellose (sample L or M, FIG. 11A) leads to small particle size withD(0.5) around 0.25 in both cases.

Example 12 Filtration

Some matrices, milling aids or facilitating agents that are used by thisinvention are not water soluble. Examples of these are microcrystallinecellulose and disintegrants such as croscarmellose and sodium starchglycolate. In order to more easily characterise the particle size of theactive after milling with these materials filtration methods can be usedto remove them allowing a characterisation of the active. In thefollowing examples naproxen was milled with lactose monohydrate andmicrocrystalline cellulose (MCC). The particle size was characterisedbefore and after filtration and the ability of the filters to letthrough the naproxen was demonstrated using HPLC assays. The millingdetails and the particle size are shown in FIG. 12 a. Note in this tablethe particle size with milling details is un-filtered. The particle sizein the rows with no milling details is after filtration. The sample thatwas filtered is indicated in the Active material section. The HPLCassays were performed by taking samples before and after filtrationthrough 10 micron poroplast filters. The samples taken were diluted togive a nominal concentration of 100 pg/ml. The HPLC assay data is shownin Table 12

Sample A was milled with 5% MCC. Before filtration the D50 was 2.5 μm,after filtration (sample B) the D50 was 183 nm. When sample B wasassayed the concentration was 94 pg/ml indicating that filtrationprocess retained little naproxen. A second milling (sample C) wasundertaken without MCC. The D50 was 160 nm as would be expected. Afterfiltration (sample D) the particle size was unchanged indicating that ifthe filtration process did remove any naproxen then it was removed in aneven way. Some of sample C was then milled with MCC for 1 minute. Thisis long enough to incorporate the MCC into the powder but not longenough to affect the particle size distribution. Two millings wereundertaken. Sample E incorporated 5% w/w MCC into the powder and SampleF 9% w/w. After incorporation of the MCC the particle size increaseddramatically. These samples where then filtered (Sample E and F) and thesize remeasured. After filtration the particle size is the same asSample C which was the starting material. The assay of samples E-Hindicates that filtration did not remove any naproxen of anysignificance. The combination of particle size and assay data clearlyshows that material such as MCC can easily and successfully be removedallowing the true particle size of the active to be measured.

Samples I and J were millings conducted with 10 and 20% w/w MCC. Theparticle size post filtration is show as sample K and L. Again thefiltration has delivered a reduction in particle size due to the removalof the MCC component. And again the HPLC assay of sample I-L showslittle naproxen was lost during filtration.

This data also demonstrates that MCC can successfully be used as comatrix in the invention disclosed herein.

TABLE 12 The HPLC assay of naproxen before and after filtration ofsamples. Sample No. HPLC Assay (μg/ml) B 94 D 93 E 99 F 96 G 98 H 97 I94 J 89 K 91 L 84

Example 13 Manufacture of Diclofenac Nanoformulation Capsules Example13(a) 18 mg

Diclofenac milled powder (666.2 g, from Example 9, Sample W) was chargedinto the bowl of a KG-5 high shear granulator. Separately, a 30% w/wsolution of povidone K30 was prepared by dissolving 60.0 g of povidoneK30 in 140.0 g of purified water. The granulator was operated at achopper speed of 250 rpm and impeller speed of 2500 rpm. A portion ofthe povidone solution (88.6 g) was introduced into the granulation overa period of approximately 9 minutes with a peristaltic pump. Anadditional 30 g of water was then added to the granulation.

The wet granules were spread on to paper-lined trays and dried in anoven at 70° C. for 2 hours. They were then manually screened through a10 mesh hand screen. After approximately 2.25 hours of drying time, theloss on drying was determined to be 0.559%.

The dried granules were processed in a Quadro CoMill fitted with a 200mesh screen and 0.225 inch spacer, run at 1265 rpm. The process yielded539.0 g of milled, dried granules.

The granules were filled into size 4 white opaque hard gelatin capsulesusing an IN-CAP® automated capsule filling machine (Dott. Bonapace & C.,Milano, Italy). The machine was set up with size 4 change parts and a 10mm dosing disc. The target fill weight was 124.8 mg, and the averageweight of an empty capsule shell was 38 mg. The machine was run at speedsetting #2. Tamping pin #4 was set to 21 mm; all other tamping pinsettings were N/A.

The filled capsules were polished in a capsule polishing machine, andthe net yield of filled capsules was 480.2 g (approximately 2,910capsules).

Example 13(b) 35 mg

Two separate granulation sublots were used for the manufacture ofDiclofenac Nanoformulation Capsules 35 mg. Granulation sublot A: 642.7 gof milled diclofenac powder (Example 9, Sample X) was charged into thebowl of a KG-5 high shear granulator. Separately, a 30% w/w solution ofpovidone K30 was prepared by dissolving 60.0 g of povidone K30 in 140.0g of purified water. The granulator was operated at an impeller speed of250 rpm and a chopper speed of 2500 rpm. A portion of the bindersolution (85.5 g) was introduced into the granulation over a period ofapproximately 8.5 minutes via a peristaltic pump. An additional 30 g ofpurified water was then added to the granulation at the same rate. Thewet granules were spread on to paper-lined trays to a thickness ofapproximately ½″.

Granulation sublot B: 519.6 g of milled diclofenac powder (Example 9,Sample Y) was charged into the bowl of a KG-5 high shear granulator.Separately, a 30% povidone solution was prepared by dissolving 60.0 g ofpovidone K30 in 140.0 g of purified water. The granulator was operatedat an impeller speed of 250 rpm and a chopper speed of 2500 rpm. Aportion of the povidone solution (69.1 g) was added to the granulationover a period of approximately 6.5 minutes. An additional 30 g of waterwas then added at the same rate. The wet granules were spread on topaper-lined trays to a thickness of approximately ½″.

The wet granules from sublots A and B were dried in an oven at 70° C.for approximately 2 hours. They were then manually screened through a 10mesh hand screen and tested for loss on drying. The LOD result was0.316%.

The dried granules were milled in a Quadro CoMill fitted with a 200 meshscreen and 0.225 inch spacer, operated at 2500 rpm. The milled granuleswere charged into an 8 qt V-blender and mixed for 5 minutes, yielding1020.2 g of granules.

The granules were filled into size 3 white opaque hard gelatin capsulesusing a MiniCap Capsule Filling Machine equipped with size 3 changeparts. The target fill weight was 242.7 mg and the average weight of anempty capsule shell was 47 mg. The granules were filled into the capsuleshells manually using a scraper. Vibration and tamping were adjusted toachieve the target fill weight. The filled capsules were polished on acapsule polishing machine, yielding 1149.2 g of filled capsules(approximately 3,922 capsules).

Example 14 Dissolution Rate of Milled Diclofenac

In this example, dissolution rate is compared between 18 mg and 35 mgnanoformulations of the invention (Example 13(a) and 13(b)), andcommercial reference diclofenac Voltarol Dispersible Tablets 50 mg(Novartis, U.K) which contain 46.5 mg of diclofenac free acid,equivalent to 50 mg of diclofenac sodium. The dissolution method usedwas Apparatus I (baskets) according to USP <711> with a stirring speedof 100 rpm. The dissolution media was 0.05% sodium lauryl sulfate andcitric acid solution buffered to pH 5.75. The dissolution volume was 900mL and dissolution medium temperature was 37° C. Samples were tested at15, 30, 45, and 60 minutes and at infinity. Infinity was defined as anadditional 15 minutes at a higher rotation speed. A sample of 1 ml wastaken at each time point, filtered and assayed by HPLC with thedetection wavelength set at 290 nm. The data in Table 14a below reportthe percent dissolved of the amount of active in each test article, forthe specified time points.

TABLE 14A Dissolution Profiles for Voltarol ® Dispersible Tablets 50 mg,Diclofenac Nanoformulation Capsules 18 mg, and DiclofenacNanoformulation Capsules 35 mg Percent Label Claim Dissolved (%)Voltarol Diclofenac Diclofenac Dispersible NanoformulationNanoformulation Time Tablets 50 mg Capsules 18 mg Capsules 35 mg 0 0 0 015 52 91 82 30 59 94.0 95 45 63 94 95 60 65 94 95 75 87 94 95

The results demonstrate that the milled diclofenac capsules dissolvemore quickly and more completely than the commercial referencediclofenac. Those of skill in the art will readily appreciate theadvantages conferred by more rapid dissolution—more active agent isavailable at any given time point. Put another way, an equal quantity ofdissolved diclofenac may be obtained with an initially smaller dosageamount of milled diclofenac, as opposed to the larger initial doserequired for the reference diclofenac to reach to the same quantity ofdissolved diclofenac. Additionally, as the results make clear, thereference diclofenac does not achieve complete dissolution even by thefinal time point, while the milled diclofenac achieves about 90%dissolution within 15 minutes. Again, a smaller dose of milleddiclofenac yields a quantity of dissolved diclofenac for which a largerdose of reference diclofenac would be required to equal.

Example 15 Bioavailability of Milled Diclofenac

This example describes a Single-Dose, Five-Way Crossover, RelativeBioavailability Study of Diclofenac Nanoformulation 18 mg and 35 mgcapsules and Cataflam® 50 mg Tablets in Healthy Subjects under Fed andFasting Conditions.

The pharmacokinetic study described in this example used DiclofenacNanoformulation Capsules 18 mg and 35 mg manufactured as described inExample 13(a) and 13(b).

Objectives:

1) To determine the relative bioavailability of diclofenac from the 35mg Test capsule versus the 50 mg Reference tablet when administered tohealthy subjects under fasting conditions.

2) To determine the effect of food on the rate and extent of absorptionof a single dose of the 35 mg Test capsule formulation of diclofenacnanoformulation administered to healthy subjects under fed and fastingconditions.

3) To determine the effect of food on the rate and extent of absorptionof a single dose of the 50 mg Reference tablet formulation of diclofenacpotassium administered to healthy subjects under fed and fastingconditions.

4) To evaluate the dose proportionality between 18 mg and 35 mg Testcapsule formulations of diclofenac nanoformulation administered tohealthy subjects under fasting conditions.

Methodology:

This was a single-center, single-dose, randomized, open-label, 5-period,5-treatment, 10-sequence crossover study that investigated the relativebioavailability and dose-proportionality of the Test product (i.e., 18mg and 35 mg nanoformulation capsules of diclofenac acid) vs. theReference product (50 mg immediate-release tablet of diclofenacpotassium [Cataflam]) administered under fed and fasting conditions.Forty (40) healthy adult male and female subjects who met all studyeligibility criteria were randomized equally on a 1:1:1:1:1 basis to oneof 10 sequences of treatment administration. Each subject received 5treatments in order of their assigned sequence according to therandomization code. Subjects entered the clinic on Day −1 of TreatmentPeriod 1 and fasted overnight. On the morning of Day 1, subjects wereadministered the Test or Reference products in the fasted state or 30minutes after the start of a FDA High-Fat Breakfast (depending on thestudy treatment). Blood samples for the pharmacokinetic (PK) evaluationof diclofenac plasma concentrations were obtained before and over 12hours following dosing. Subjects were then discharged and returned tothe clinic after a 7-day washout interval to continue the treatmentsequence for Periods 2, 3, 4, and 5. A blood sample for safetyassessments was collected with the last PK sample in Treatment Period 5.Adverse event (AE) information elicited during confinement or reportedat outpatient visits was reviewed and documented.

Number of Subjects (Planned and Analyzed):

Number of subjects planned for enrollment: up to 40

Number of subjects enrolled in study: 40

Number of subjects completing study: 38

Number of subjects bioanalytically analyzed: 30

Number of subjects statistically analyzed: 30

Diagnosis and Main Criteria for Inclusion:

Subjects were males and females who provided written informed consent,were at least 18 years of age, and had a body weight of at least 110pounds and a body mass index (BMI) between 18 and 30 kg/m², and werehealthy on the basis of medical history, physical examination,electrocardiogram (ECG), and clinical laboratory test results. Allfemales were non-pregnant and non-nursing; females of child-bearingpotential agreed to take precautions to prevent pregnancy. Eligibilitycriteria required that subjects demonstrate negative test results forhepatitis B, hepatitis C, and human immunodeficiency virus, as well as anegative urine screen for drugs of abuse and breathalyzer test foralcohol.

Test Product, Dose, and Mode of Administration:

The Test products were diclofenac acid nanoformulation 18 mg and 35 mgcapsules.

The 18 mg Test product was administered as Treatment A. Subjectsassigned to Treatment A received a single 18 mg capsule by mouth with240 mL of water after an overnight fast.

The 35 mg Test product was administered as Treatments B and C. Subjectsassigned to Treatment B received a single 35 mg capsule by mouth with240 mL of water after an overnight fast. Subjects assigned to TreatmentC received a single 35 mg capsule by mouth with 240 mL of water 30minutes after the start of a FDA High-Fat Breakfast.

Duration of Treatment:

The duration of treatment was a single dose in each period.

Reference Therapy, Mode of Administration, and Lot Number:

The Reference product was Cataflam (diclofenac potassium) 50 mg tablets,manufactured by Patheon Inc, Whitby Operations and distributed byNovartis Pharmaceutical Corporation. A single lot of the Referenceproduct was used in this study (Lot number C7C02722). The Referenceproduct was administered as Treatments D and E. Subjects assigned toTreatment D received a single 50 mg tablet by mouth with 240 mL of waterafter an overnight fast. Subjects assigned to Treatment E received asingle 50 mg tablet by mouth with 240 mL of water 30 minutes after thestart of a FDA High-Fat Breakfast.

Criteria for Evaluation:

Pharmacokinetic:

Blood samples for measurement of diclofenac concentrations in plasmawere collected pre-dose and 0.083, 0.167, 0.25, 0.33, 0.50, 0.67, 1,1.33, 1.67, 2, 2.33, 2.67, 3, 3.67, 4, 4.5, 5, 6, 8, 10, and 12 hourspost-dose. Primary PK variables included: area under theconcentration-time curve from time zero to the time of the last samplewith a quantifiable concentration (AUC_(0-t)); area under theconcentration time curve from time zero extrapolated to infinity(AUC_(0-∞)); and, measured maximal concentrations (C_(max)). SecondaryPK variables included: time to reach maximum concentration (T_(max));terminal elimination rate constant (K_(e)); and terminal eliminationhalf-life (T_(1/2)).

Safety:

A physical examination, serology test for HIV, Hepatitis B, andHepatitis C, as well as a urine drug screen were performed at theScreening Visit. Samples for general clinical laboratory tests werecollected, 12-lead ECG tracings were obtained, vital signs weremeasured, and pregnancy tests (for female subjects) were performed atthe Screening Visit and at specified time points. During the study,subjects were monitored for clinical and laboratory evidence of adverseevents.

Statistical Methods:

Pharmacokinetic:

Statistical analyses were performed using the mixed model procedure ofthe SAS® statistical program (PC Version 9.1.3) in a Windows XPProfessional environment. The pharmacokinetic parameter estimates wereevaluated using mixed model analyses (PROC MIXED). The model includedfixed effects for sequence, period, and treatment; and random effectsfor subject nested within sequence. The least-squares means and the meanstandard error values from these analyses were used to construct the 90%confidence intervals for the relative bioavailability evaluationsaccording to the FDA's recommended procedures. The dose normalized AUCand C_(max) values for the 18 mg and 35 mg Test products wereIn-transformed and compared by Analysis of Variance (ANOVA). Asspecified in the protocol, dose proportionality was to be concluded ifthe overall treatment effect was not significant at the 5% level, or ifthe 90% confidence intervals for the ratios of geometric means containedthe value “1.00”.

Safety:

Adverse events were coded using the Medical Dictionary for RegulatoryActivities (MedDRA) and listed by system organ class (SOC) and preferredterm (PT). Treatment-emergent AEs were summarized by incidence,relatedness to study drug, and severity.

Summary—Results

Demographic Characteristics of Subjects:

Forty (40) subjects were randomized into treatment and 38 subjects (95%)completed all 5 study periods. Two subjects voluntarily withdrew consentprior to dosing in Period 2. The 40 subjects who received at least onedose of study drug were included in the full analysis set and ranged inage from 21 to 56 years, with a mean age of 34.6 years. There were 27male subjects (67.5%) and 13 female subjects (32.5%). With regard torace/ethnicity, 32 subjects (80.0%) were Black, 6 subjects (15.0%) wereCaucasian, and 2 subjects (5.0%) were Hispanic. The mean height was172.9 cm, with a range of 151 to 189 cm. The mean body weight was 77.4kg, with a range of 52.9 to 104.8 kg. The mean BMI was 25.8 kg/m², witha range of 20.0 to 29.7 kg/m². Demographic findings were reflective of ahealthy adult population.

Pharmacokinetic Results:

Consistent with the Statistical Analysis Plan, all available data fromthe first 30 of the 38 subjects who completed all 5 periods were used inthe pharmacokinetic analyses. Statistical test results onpharmacokinetic parameters for diclofenac are summarized in the Tables15a-d below.

TABLE 15a Treatments B:D (35 mg Test product vs. 50 mg Reference product[fasting conditions]) 35 mg Test product vs. 50 mg Reference product -Fasted subjects Test Pro- Reference Pharmacokinetic duct 35 Product 50Parameter/Unit mg Fasted^(a) mg Fasted^(a) Ratio^(b) 90% CI^(c)AUC_((0-t)) hr*ng/mL 1132 1432 0.791* 0.758,0.825 AUC_((0-∞)) hr*ng/mL1152 1449 0.795* 0.764,0.828 C_(max) ng/mL 1179 1268 0.930* 0.789,1.096T_(max) ^(d) hr 0.559 0.737 0.758 — (0.500) (0.667) K_(e) 1/hr 0.39770.3763 1.057 — T_(1/2) hr 1.83 1.92 0.956 —

AUC_((0-t)) and AUC_((0-∞)) values for the Test (35 mg capsule) productwere approximately 20% lower than observed for the Reference product (50mg tablet). C_(max) for the Reference product was only 7% greater thanthe Test product and was not statistically significant (α=0.05). The 35mg Test capsules and the 50 mg Reference tablets (Cataflam) were notbioequivalent when given to fasted subjects.

Abbreviations: ANOVA (analysis of variance); AUC_((0-t)) (area under theconcentration-time curve from zero to the last measurableconcentration); AUC_((0-∞)) (area under the concentration-time curvefrom zero to infinity); CI (confidence interval); C_(max) (measuredmaximal plasma concentration); K_(e) (terminal elimination rateconstant); T_(1/2) (terminal elimination half life); T_(max) (time toachieve maximum concentration).

a. Least-squares geometric means for areas and peak concentrations.Least squares arithmetic means for other parameters.

b. Ratio calculated as Test Fasted least-squares mean divided by theReference Fasted least-squares mean.

c. Confidence interval on the Test-to-Reference ratio.

d. Mean (median) reported for T_(max).

* Comparisons were detected as statistically significant with α=0.05.

TABLE 15b Treatments C:B (35 mg Test product [fed vs. fasted subjects])35 mg Test product - Fed versus Fasted Subjects Pharmacokinetic TestProduct Parameter/Unit 35 mg Fed^(a) 35 mg Fasted^(a) Ratio^(b) 90%CI^(c) AUC_((0-t)) hr*ng/mL 1034 1132 0.913* 0.876, 0.952 AUC_((0-∞))hr*ng/mL 1073 1152 0.931* 0.893, 0.970 C_(max) ng/mL 490 1179 0.416*0.353, 0.489 T_(max) ^(d) hr 1.93 (1.67) 0.559 (0.500) 3.445* — K_(e)1/hr 0.3275 0.3977 0.824* — T_(1/2) hr 2.26 1.83 1.234* —

Food reduced AUC_((0-t)) and AUC_((0-∞)) values by 9% and 7%,respectively. C_(max) decreased by 58%. All parameters werestatistically significant (α=0.05) indicating a food effect for the 35mg Test product.

Abbreviations: ANOVA (analysis of variance); AUC_((0-t)) (area under theconcentration-time curve from zero to the last measurableconcentration); AUC_((0-∞)) (area under the concentration-time curvefrom zero to infinity); CI (confidence interval); C_(max) (measuredmaximal plasma concentration); K_(e) (terminal elimination rateconstant); T_(1/2) (terminal elimination half life); T_(max) (time toachieve maximum concentration).

a. Least-squares geometric means for areas and peak concentrations.Least squares arithmetic means for other parameters.

b. Ratio calculated as Test Fed least-squares mean divided by the TestFasted least-squares mean.

c. Confidence interval on the Test Fed-to-Test Fasted ratio.

d. Mean (median) reported for T_(max).

* Comparisons were detected as statistically significant by ANOVA, withα=0.05.

TABLE 15c Treatments E:D (50 mg Reference product [fed vs. fastedSubjects]) 50 mg Reference Product - Fed versus Fasted SubjectsPharmacokinetic Reference Product Parameter/Unit 50 mg Fed^(a) 50 mgFasted^(a) Ratio^(b) 90% CI^(c) AUC_((0-t)) hr*ng/mL 1308 1432 0.914*0.877,0.952 AUC_((0-∞)) hr*ng/mL 1334 1449 0.920* 0.884,0.958 C_(max)ng/mL 922 1268 0.728* 0.619,0.856 T_(max) ^(d) hr 1.70 0.737 2.299* —(1.33) (0.667) K_(e) 1/hr 0.3424 0.3763 0.910 — T_(1/2) hr 2.17 1.921.134* —

Food reduced AUC_((0-t)) and AUC_((0-∞)) by 9% and 8%, respectively.C_(max) decreased by 27%. These PK parameters were statisticallysignificant (α=0.05), indicating a food effect for the Referenceproduct.

Abbreviations: ANOVA (analysis of variance); AUC_((0-t)) (area under theconcentration-time curve from zero to the last measurableconcentration); AUC_((0-∞)) (area under the concentration-time curvefrom zero to infinity); CI (confidence interval); C_(max) (measuredmaximal plasma concentration); K_(e) (terminal elimination rateconstant); T_(1/2) (terminal elimination half life); T_(max) (time toachieve maximum concentration).

a. Least-squares geometric means for areas and peak concentrations.Least squares arithmetic means for other parameters.

b. Ratio calculated as Reference Fed least-squares mean divided by theReference Fasted least-squares mean.

c. Confidence interval on the Reference Fed-to-Reference Fasted ratio.

d. Mean (median) reported for T_(max).

* Comparisons were detected as statistically significant by ANOVA, withα=0.05.

TABLE 15d Dose proportionality for Treatments A:B (18 mg vs. 35 mg TestProduct [Fasted Subjects]) 18 mg Test Product versus 35 mg TestProduct - Fasted Subjects Pharmacokinetic Test Product Parameter/Unit 18mg Fasted^(a) 35 mg Fasted^(a) Ratio^(b) 90% CI^(c) AUC_((0,t)) hr*ng/mL1061 1132 0.938* 0.899, 0.977 AUC_((0-∞)) hr*ng/mL 1090 1152 0.946*0.909, 0.984 C_(max) ng/mL 1174 1179 0.996* 0.847, 01.172 T_(max) ^(d)hr 0.571 0.559 1.022 — (0.500) (0.500) K_(e) 1/hr 0.4760 0.3977 1.197* —T_(1/2) hr 1.50 1.83 0.821* —

Dose-normalized In-transformed AUC_((0-t)) and AUC_((0-∞)) differenceswere statistically significant (α<0.05), and the 90% confidenceintervals for their ratios did not contain the value “1.00”, thus theseparameters did not pass the per protocol dose-proportionality test.Alternatively, dose-proportionality may still be concluded for the 18 mgand 35 mg Test products since (1) the 90% confidence intervals of theratios of the dose-normalized In-transformed C_(max), AUC, and AUC weretotally contained within the 0.800 and 1.250 acceptance range forbioequivalence; and (2) the ratios of dose-normalized geometric meansbetween the 18 mg and 35 mg Test capsules were 0.996, 0.938, and 0.946,respectively for C_(max), AUC_((0-t)), and AUC_((0-∞)).

Abbreviations: ANOVA (analysis of variance); AUC_((0-t)) (area under theconcentration-time curve from zero to the last measurableconcentration); AUC_((0-∞)) (area under the concentration-time curvefrom zero to infinity); CI (confidence interval); C_(max) (measuredmaximal plasma concentration); K_(e) (terminal elimination rateconstant); T_(1/2) (terminal elimination half life); T_(max) (time toachieve maximum concentration).

a. Least-squares geometric means for areas and peak concentrations.Least squares arithmetic means for other parameters. AUC_((0-t)),AUC_((0-∞)), and C_(max) for the 18 mg were dose-normalized bymultiplying the ratio of 35 mg divided by 18 mg (equal to 1.944).

b. Ratio calculated as dose-normalized least-squares mean for the 18 mgTest product divided by the least-squares mean for the 35 mg Testproduct.

c. Confidence interval on the dose-normalized 18 mg Test-to-35 mg Testratio.

d. Mean (median) reported for T_(max).

* Comparisons were detected as statistically significant by ANOVA, withα=0.05.

Safety Results:

A total of 40 (100%) subjects were included in the safety population.Thirteen (13) treatment-emergent adverse events (AEs) were experiencedby 7 subjects (18.0%). Six (6) treatment-emergent AEs were reported by 5subjects (12.5%) who received the Test product and 7 treatment-emergentAEs were reported by 2 subjects (5.0%) who received the Referenceproduct. Ten (10) of the 13 treatment-emergent AEs (76.9%) wereconsidered to be mild in severity; 3 (23.0%) were considered to bemoderate (i.e., vomiting and headache [Subject 15]) and none wereserious or life-threatening. Fatigue was the treatment-emergent AD withthe highest incidence overall, i.e., reported by 3 subjects (8.0%) whoreceived the Test product (diclofenac nanoformulation 35 mg capsule).Six (6) out of 40 subjects (15.0%) reported 9 treatment-emergent AEsthat were determined to be at least possibly related to study drugadministration. No clinically significant changes in laboratory resultsor vital signs occurred, in the opinion of the Investigator. There wereno deaths or other serious adverse events in this study.

Conclusions:

This was a single-center, single-dose, randomized, open-label, 5-period,5-treatment 10-sequence crossover study that investigated the relativebioavailability and dose-proportionality of the Test product (i.e., 18mg and 35 mg nanoformulation capsules of diclofenac acid) vs. theReference product (50 mg immediate-release tablet of diclofenacpotassium [Cataflam]) administered under fed and fasting conditions.Forty (40) healthy male and female adults were enrolled in the study.Eligible subjects received 5 treatments in order of their assignedsequence according to the randomization schedule. There was a 7-daywashout interval between each Treatment Period. Thirty-eight (38; 95%)subjects completed all 5 study periods. Two 92; 5%) subjects voluntarilywithdrew prior to dosing in Period 2. Pharmacokinetic blood samples wereobtained pre-dose and over 12 hours following dosing. During the study,subjects were monitored for clinical and laboratory evidence of adverseevents.

A single dose of diclofenac (18 mg, 35 mg, or 50 mg) was safe andwell-tolerated. Seven subjects (7; 18.0%) reported a total of 13treatment-emergent AEs. Six (6; 15%) subjects reported 9treatment-emergent AEs that were determined to be at least possiblyrelated to study drug administration. All treatment-emergent AEs wereconsidered to be mild in severity, except for vomiting and 2 occurrencesof headache experienced by Subject 15. No deaths or serious adverseevents occurred.

All available data from the first 30 subjects who completed all 5periods of the study were used in the pharmacokinetic analyses.Conclusions based on statistical testing of the PK parameters fordiclofenac are summarized below.

35 mg Test Product Vs. 50 mg Reference Product (Fasted Subjects):

These two formulations were not bioequivalent under fasting conditionssince the 90% confidence intervals on the geometric mean and peakconcentration ratios for diclofenac were outside the interval 0.800 to1.250.

mg Test Product (Fed Vs. Fasted Subjects):

Food decreased AUC_((0-t)) and AUC_((0-∞)) values by 9% and 7%respectively. C_(max) was decreased by 58%. Conversely, T_(max) was 22minutes slower when the 35 mg Test product was administered under fedconditions.

50 mg Reference Product (Fed Vs. Fasted Subjects):

Food decreased AUC_((0-t)) and AUC_((0-∞)) values by 9% and 8%respectively. C_(max) was decreased by 27%. Conversely, T_(max) was 58minutes slower when the 50 mg Reference product was administered underfed conditions.

35 mg Test Product Vs. 50 mg Reference Product (Fed Subjects):

These two formulations were not bioequivalent under fed conditions sincethe 90% confidence intervals on the geometric mean and peakconcentration ratios for diclofenac were outside the interval 0.800 to1.250.

Dose Proportionality Evaluation (18 mg Vs. 35 mg Test Product in FastedSubjects):

Although the 18 mg and 35 mg Test products were not identified asdose-proportional based on the per protocol analysis,dose-proportionality may still be concluded since: (1) the 90%confidence intervals for the ratios of dose-normalized In-transformedC_(max), AUC_((0-t)), and AUC_((0-∞)) were totally contained within the0.800 and 1.250 acceptance range for bioequivalence; and (2) the ratiosof dose-normalized geometric means between the 18 mg and 35 mg Testcapsules were 0.996, 0.938, and 0.946 respectively, for C_(max),AUC_((0-t)), and AUC_((0-∞)).

Example 16 Efficacy and Safety of Milled Diclofenac

This Example describes a Phase 2, Phase 2, Randomized, Double-Blind,Single-Dose, Parallel-Group, Active- and Placebo-Controlled Study ofDiclofenac Nanoformulation Capsules for the Treatment of Pain AfterSurgical Removal of Impacted Third Molars.

The phase II efficacy study described in this example uses DiclofenacNanoformulation Capsules 18 mg and 35 mg manufactured as described inExample 13(a) and (b).

Objectives:

The primary objective of this study is to evaluate the analgesicefficacy and safety of Diclofenac Nanoformulation Capsules compared withplacebo in subjects with acute dental pain after third molar extraction.The secondary objective of this study is to evaluate the time to onsetof analgesia for Diclofenac Nanoformulation Capsules compared with thestandard formulation of celecoxib.

Number of Subjects:

Planned enrollment (and/or completion): Approximately 200 subjects (50in each treatment group) will be enrolled.

Subject Population:

Inclusion Criteria:

A subject will be eligible for study entry if all of the followinginclusion criteria are met:

-   1. Is male or female ≧18 and ≦50 years of age.-   2. Requires extraction of 2 or more third molars. At least 1 of the    third molars must be a fully or partially bone-impacted mandibular    molar. If only 2 molars are removed, then they must be ipsilateral.-   3. Experiences moderate to severe pain intensity within 6 hours    after surgery, as measured by a Visual Analog Scale (VAS) score of    ≧50 mm on a 100-mm scale.-   4. Has a body weight of ≧45 kg and a body mass index (BMI)≦35 kg/m².-   5. If female and of childbearing potential, is nonlactating and    nonpregnant (has negative pregnancy test results at screening    [serum] and on the day of surgery prior to surgery [urine]).-   6. If female, is either not of childbearing potential (defined as    postmenopausal for at least 1 year or surgically sterile [bilateral    tubal ligation, bilateral oophorectomy, or hysterectomy]) or    practicing 1 of the following medically acceptable methods of birth    control:    -   a. Hormonal methods such as oral, implantable, injectable, or        transdermal contraceptives for a minimum of 1 full cycle (based        on the subject's usual menstrual cycle period) before the study        drug administration.    -   b. Total abstinence from sexual intercourse (since the last        menses before study drug administration).    -   c. Intrauterine device (IUD).    -   d. Double-barrier method (condoms sponge, diaphragm, or vaginal        ring with spermicidal jellies or cream).-   7. Is in good health, in the opinion of the investigator.-   8. Is able to provide written informed consent to participate in the    study and able to understand the procedures and study requirements.-   9. Must voluntarily sign and date an informed consent form (ICF)    that is approved by an Institutional Review Board (IRB) prior to the    conduct of any study procedure.-   10. Is willing and able to comply with study requirements (including    diet and smoking restrictions), complete the pain evaluations,    remain at the study site overnight, and return for follow-up 7±2    days after surgery.    Exclusion Criteria:

A subject will not be eligible for study entry if any of the followingexclusion criteria are met:

-   -   1. Has a known history of allergic reaction or clinically        significant intolerance to acetaminophen, aspirin, or any        nonsteroidal anti-inflammatory drug (NSAIDs, including        diclofenac and celecoxib); history of NSAID-induced bronchospasm        (subjects with the triad of asthma, nasal polyps, and chronic        rhinitis are at greater risk for bronchospasm and should be        considered carefully); or hypersensitivity, allergy, or        significant reaction to sulfa (including sulfonamide) medicines,        ingredients of the study drug, or any other drugs used in the        study including anesthetics and antibiotics that may be required        on the day of surgery.    -   2. Has tested positive either on the urine drug screen or on the        alcohol breathalyzer test. Subjects who test positive at        screening only and can produce a prescription for the medication        from their physician may be considered for study enrollment at        the discretion of the investigator.    -   3. Has known or suspected history of alcoholism or drug abuse or        misuse within 2 years of screening or evidence of tolerance or        physical dependence before dosing with the study drug.    -   4. Has received or will require any medication (except hormonal        contraceptives, vitamins, or nutritional supplements) within 5        half-lives (or, if half-life is unknown, within 48 hours) before        dosing with study drug.    -   5. Has any clinically significant unstable cardiac, respiratory,        neurological, immunological, hematological, or renal disease or        any other condition that, in the opinion of the investigator,        could compromise the subject's welfare, ability to communicate        with the study staff, or otherwise contraindicate study        participation.    -   6. Has a history or current diagnosis of a significant        psychiatric disorder that, in the opinion of the investigator,        would affect the subject's ability to comply with the study        requirements.    -   7. Is receiving systemic chemotherapy, has an active malignancy        of any type, or has been diagnosed with cancer with 5 years of        screening (excluding squamous or basal cell carcinoma of the        skin).    -   8. Has a history of clinically significant (investigator        opinion) gastrointestinal (GI) event within 6 months before        screening or has any history of peptic or gastric ulcers or GI        bleeding.    -   9. Has a surgical or medical condition of the GI or renal system        that might significantly alter the absorption, distribution, or        excretion of any drug substance.    -   10. Is considered by the investigator, for any reason        (including, but not limited to, the risks described as        precautions, warnings, and contraindications in the current        version of the Investigator's Brochure [IB] for Diclofenac        Nanoformulation Capsules), to be an unsuitable candidate to        receive the study drug.    -   11. Has history of chronic use (defined as daily use for >2        weeks) of NSAIDs, opiates, or glucocorticoids (except inhaled        nasal steroids and topical corticosteroids), for any condition        within 6 months before dosing with study drug. Aspirin at a        daily dose of 325 mg is allowed for cardiovascular (CV)        prophylaxis if the subject has been on a stable dose regimen for        ≧30 days before screening and has not experienced any relevant        medical problem.    -   12. Has a significant renal or hepatic disease, as indicated by        the clinical laboratory assessment (results ≧3 times the upper        limit of normal [ULN] for any liver function test, including        aspartate aminotransferase [AST], alanine aminotransferase        [ALT], and lactate dehydrogenase, or creatinine ≧1.5 times the        ULN) or has any clinically significant laboratory findings at        screening that in the investigator's opinion contraindicate        study participation.    -   13. Has significant difficulties swallowing capsules or is        unable to tolerate oral medication.    -   14. Previously participated in another study of Diclofenac        Nanoformulation Capsules, or received any investigational drug        or device or investigational therapy within 30 days before        screening.        Design:

This is a phase 2, multicenter, randomized, double-blind, single-dose,parallel-group, active- and placebo-controlled study to evaluate theefficacy and safety of Diclofenac Nanoformulation Capsules (18 mg and 35mg) in subjects with postoperative dental pain. Eligible subjects willcomplete all screening procedures within 28 days before the surgery.

At screening, subjects will provide written informed consent toparticipate in the study before any protocol-specified procedures orassessments are completed. On Day 1, subjects who continue to beeligible for study participation after completing screening proceduresand assessments are will undergo extraction of 2 or more third molars.At least 1 of the third molars must be a fully or partiallybond-impacted mandibular molar. If only 2 molars are removed, then theymust be ipsilateral. All subjects will receive local anesthesia (2%lidocaine with 1:100,000 epinephrine). Nitrous oxide will be allowed atthe discretion of the investigator. Subjects who experience moderate tosevere pain intensity (a score of 50 mm on a 100-mm VAS) within 6 hoursafter surgery and who continue to meet all study entry criteria will berandomized in a 1:1:1:1 ratio to receive 1 oral dose of DiclofenacNanoformulation Capsules (18 mg or 35 mg), celecoxib capsules (400 mg),or placebo. Study drug will be administered by an unblended, third-partydoser who will not conduct any efficacy or safety assessments.

Subjects will assess their baseline pain intensity (VAS) beforereceiving study drug (predose, Time 0) and their pain intensity (VAS)and pain relief (5-point categorical scale) at the following timepoints: 15, 30, and 45 minutes, and 1, 1.5, 2, 3, 4, 5, 6, 7, 8, 10, and12 hours after Time 0; and immediately before the first dose of rescuemedication. The 2-stopwatch method will be used to record the time toperceptible and time to meaningful pain relief, respectively. Subjectswill complete a global evaluation of study drug 12 hours after Time 0 orimmediately before the first dose of rescue medication (whichever occursfirst). Vital signs will be recorded after the subject has been in asitting position for 5 minutes at the following times: before surgery,before Time 0, 12 hours after Time 0, and/or immediately before thefirst dose of rescue medication. Adverse events (AEs) will be monitoredand recorded from the time of signing of the ICF until the Follow-upVisit (or Early Termination Visit). During the 12 hours following Time0, subjects will complete efficacy and safety assessments. Subjects willremain at the study site overnight and will be discharged the morning ofDay 2. Upon discharge from the study site, subjects will be given adiary to record concomitant medications taken and AEs experienced afterdischarge.

Acetaminophen (1000 mg) will be permitted as the initial rescuemedication. Subjects will be encouraged to wait at least 60 minutesafter receiving study drug before taking rescue medication. Additionalanalgesic rescue medication may be administered at the discretion of theinvestigator if the protocol-specified rescue medication is deemedinadequate. Subjects are not permitted to take medications (excepthormonal contraceptives, vitamins, nutritional supplements, and studydrug) within 5 half-lives (or, if half-life is unknown, within 48 hours)before dosing with study drug until discharge from the study (Day 2).Other restrictions include the following: alcohol use is prohibited from24 hours before surgery until discharge on Day 2; nothing by mouth (NPO)from midnight before surgery until 1 hour after surgery; clear liquidsonly are allowed starting 1 hour after surgery until 1 hour afterdosing; 1 hour after dosing diet may be advanced according to standardpractice.

Upon discharge from the study site, subjects may be prescribed painmedication for use at home according to the standard practice of thestudy site. On Day 8 (±2 days), subjects will return to the study sitefor an abbreviated confirmatory physical assessment and concomitantmedication and AE assessments.

Study Drug:

Diclofenac Nanoformulation Capsules (18 mg and 35 mg) for oraladministration

Reference Products:

Celecoxib 200-mg capsules administered as a 400-mg dose

Placebo

Treatment Regimens

Eligible subjects meeting all study entry criteria will be randomized toreceive 1 of the following treatments:

One 18-mg Diclofenac Nanoformulation Capsule and 1 placebo capsule

One 35-mg Diclofenac Nanoformulation Capsule and 1 placebo capsule

Two 200-mg celecoxib capsules

Two placebo capsules

Study Duration:

Up to approximately 5 weeks for each subject, including a 4-weekscreening period and a posttreatment Follow-up Visit approximately 1week after dosing with study drug.

Investigative Sites or Countries:

Two study sites in the United States (US).

Study Endpoints:

Efficacy Endpoints:

The primary efficacy endpoint is the sum of total pain relief (TOTPAR)over 0 to 12 hours (TOTPAR-12) after Time 0.

The secondary endpoints are the following:

-   -   TOTPAR over 0 to 4 hours (TOTPAR-4) and over 0 to 8 hours        (TOTPAR-8) after Time 0.    -   VAS pain intensity difference (VASPID) at each scheduled time        point after Time 0.    -   Time to onset of analgesia (measured as time to perceptible pain        relief confirmed by meaningful pain relief).    -   VAS pain intensity score at each scheduled time point.    -   VAS summed pain intensity difference (VASSPID) over 0 to 4 hours        (VASSPID-4), over 0 to 8 hours (VASSPID-8), and over 0 to 12        hours (VASSPID-12) after Time 0.    -   Summed pain relief and intensity difference (sum of TOTPAR and        VASSPID [SPRID]) over 0 to 4 hours (SPRID-4), over 0 to 8 hours        (SPRID-8), and over 0 to 12 hours (SPRID-12) after Time 0.    -   Pain relief score at each scheduled time point after Time 0.    -   Peak pain relief.    -   Time to peak pain relief.    -   Time to first perceptible pain relief.    -   Time to meaningful pain relief.    -   Proportion of subjects using rescue medication.    -   Time to first use of rescue medication (duration of analgesia).    -   Patient's global evaluation of study drug.        Safety Endpoints:

The safety endpoints are the incidence of treatment-emergent AEs (TEAEs)and changes in vital sign measurements.

Statistical Methods Summary:

Analysis Populations:

The analysis populations include the following:

-   -   The intent-to-treat (ITT) population will consist of all        subjects who are treated with study drug and who have at least 1        pain relief assessment after Time 0. The ITT population is the        primary population for the efficacy analysis.    -   The per-protocol (PP) population will consist of all ITT        subjects who remain in the study for at least 12 hours of        treatment and who do not incur a major protocol violation that        would challenge the validity of their data. This population will        be utilized to evaluate the sensitivity of the primary efficacy        analysis.    -   The safety population will include all subjects who are treated        with study drug. The safety population is the population for all        safety assessments.        Subject Characteristics:

Demographic and baseline characteristics (including age, sec, race,weight, height, BMI, medical history, surgery duration, and baselinevalues of efficacy variables) will be summarized for each treatmentgroup and for the overall population by descriptive statistics. Noformal statistical analyses will be performed.

Efficacy Analyses:

The null hypothesis in this study is that TOTPAR-12 for placebo is equalto TOTPAR-12 for 35-mg Diclofenac Nanoformulation Capsules. It will beanalyzed using analysis of covariance (ANCOVA) models, which includetreatment effect and significant covariates. The effect of potentialcovariates, such as sex, baseline pain intensity, and surgical traumarating, will be assessed using appropriate ANCOVA models. The analysiswill be based on a 2-sided test as the significance level of 0.05.

Other comparisons between the treatment regimens, including 18-mgDiclofenac Nanoformulation Capsules versus placebo and 400-mg celecoxibcapsules versus placebo will be considered secondary. No P valueadjustment will be made for multiple endpoints or multiple comparisons.Each efficacy endpoint will be summarized descriptively by treatmentgroup.

For continuous secondary endpoints such as pain intensity score, VASPIDat each scheduled time point, peak pain intensity, TOTPAR-4, TOTPAR-8,VASSPID-4, VASSPID-8, VASSPID-12, SPRID-4, SPRID-8, and SPRID-12,descriptive statistics (such as mean, standard error, median, minimum,and maximum) will be provided for each treatment regimen. Nominal Pvalues from 2-sample tests comparing the placebo group with othertreatment groups will be provided, but no formal statistical inferenceswill be drawn on the basis of these tests.

For ordinal secondary endpoints, such as pain relief at each scheduledtime point, peak pain relief, and global evaluation of study drug,descriptive summaries will be provided to include the number andpercentage of subjects within each category for each treatment group.Nominal P values from Fisher's exact tests (or chi-square tests, asappropriate) comparing the placebo group with other treatment groupswill be provided, but no formal statistical inferences will be drawn onthe basis of these tests.

For each time-to-event endpoint, the Kaplan-Meier method will be used toevaluate the treatment effect. Time to onset of analgesia (measured astime to perceptible pain relief confirmed by meaningful pain relief)will be based on data collected using the 2-stopwatch method. Time toonset of analgesia will be right-censored at 12 hours for subjects whodo not experience both perceptible pain relief and meaningful painrelief during the 12-hour interval after Time 0. The summary table willprovide the number of subjects analyzed, the number of subjectscensored, estimates for the quartiles, and 95% confidence intervals(Cis) for the estimated median and the restricted mean estimate. Pvalues form the Wilcoxon or log-rank tests (as appropriate) will also beused to examine treatment effect. Cox proportional hazard models will beused to explore such potential covariates as sex, baseline painintensity, and surgical trauma rating, if appropriate.

For the proportion of subjects using rescue medication, a logisticregression model that adjusts for baseline pain intensity, ifappropriate, will be used to evaluate the treatment effect. Subgroupanalysis by sex may be performed if it is confirmed to be astatistically significant covariate for TOTPAR-12. Baseline values aredefined as the last measurements taken before dosing with a study drug.

For pain intensity, missing observations will be imputed usingbaseline-observation-carried-forward (BOCF) for subjects who withdrawfrom the study due to lack of efficacy or an AE/intolerance to studydrug. The BOCF imputation will be applied in place of all scheduledassessments after the time of early termination due to lack of efficacyor an AE/intolerance to study drug using the baseline observation takenbefore Time 0.

For subjects who withdraw from the study due to reasons other than lackof efficacy or an AE/intolerance to study drug, missing observations forpain intensity and pain relief will be imputed usinglast-observation-carried-forward (LOCF). The LOCF imputation will beapplied in place of all scheduled assessments after the time of earlytermination due to reasons other than lack of efficacy or anAE/intolerance to the drug.

For subjects who take any dose of rescue medication, subsequent measuresafter the first dose of rescue medication will be disregarded. Instead,all scheduled assessments after the first dose of rescue medication willbe imputed using BOCF using the baseline observation taken before Time0. Single missing data points will be imputed using linearinterpolation, if they do not occur at the end of the study. For otherconditions before early termination or rescue medication, missing datawill be imputed using LOCF.

Safety Analysis:

Data listings will be provided for protocol-specified safety data. TheMedical Dictionary for Regulatory Activities (MedDRA) (Version 9.1 orhigher) will be used to classify all AEs with respect to system organclass and preferred term. Adverse event summaries will include onlyTEAEs, which will be summarized for each treatment group. Fisher's2-sided exact test will be used to compare the rates of occurrencebetween the placebo and Diclofenac Nanoformulation Capsule groups forall TEAEs.

For vital sign measurements, descriptive statistics will be provided ateach scheduled time point for each treatment group. Changes fromBaseline for vital signs will be calculated for each subject, anddescriptive statistics will be provided on changes in vital signs fromBaseline for each treatment group at each scheduled time point afterBaseline. No formal statistical tests will be performed.

Sample Size:

The standard deviation of TOTPAR-12 is assumed to be 14.0. A sample sizeof 50 subjects per treatment group will provide 80% power to detect aminimal difference of 8.0 in TOTPAR-12 using a 2-sample t-test with a0.05 two-sided significance level (nQuery v6.0).

TABLE 16a Schedule of Events B^(a) D A C E F G H I J K Written InformedConsent X Assign a screening number X Inclusion/exclusion criteria X XDemographics X Medical History X X^(b) Physical Examination^(c) X XVital signs^(d) X X X X X Height, weight, and BMI X Clinical laboratorytests X (hematology, chemistry, urinalysis) Pregnancy test for female XX subjects of childbearing potential^(c) Urine drug screen X X Alcoholbreathalyzer test X Oral radiography^(f) X Review study restrictions Xwith subject Pain intensity (VAS)^(g) X X X X Randomization X Dosingwith study drug X Stopwatch assessment^(h) X Pain relief (5-point X X Xcategorical scale)^(g) Global evaluation of study X drug^(i) Concomitantmedications X^(b) X X X X X X X Adverse events^(j) X X X X X X X XDispense rescue medication/ X pain medications Collect unused rescue Xmedication/pain medications Dispense/collect subject diary X X Dischargefrom study site X A: Screening (Days −28 to −1); B: Day of Surgery (Day1); C: Preop; D: Postop; E: Predose; F: 0 h; G: 15, 30, 45 min; H: 1,1.5, 2, 3, 4, 5, 6, 7, 8, 10 h; I: 12 h; J: Day 2; K: Follow-up (Day 8 ±2 days or ET). Abbreviations: BMI, body mass index; ET, earlytermination; h, hour; min, minute; preop, preoperative; postop,postoperative; VAS, Visual Analogue Scale. ^(a)Times listed are relativeto dosing with study drug. ^(b)Medical history and concomitantmedication use since screening will be updated on Day 1 before surgery.^(c)A complete physical examination (excluding the genitourinaryexamination) will be performed at screening. An abbreviated confirmatoryphysical assessment, including an examination of the subject's mouth andneck, will be performed at the Follow-Up visit (or Early Terminationvisit) ^(d)Vital signs will be recorded after the subject has been in asitting position for 5 minutes at the following times: at screening,before surgery, before Time 0, 12 hours after Time 0, and/or immediatelybefore the first dose of rescue medication, and the Follow-up Visit (orEarly Termination visit). ^(e)Serum pregnancy test at screening andurine pregnancy test before surgery on Day 1 (female subjects ofchildbearing potential only). Test results must be negative for thesubject to continue in the study. ^(f)Oral radiographs taken within 1year before screening will be acceptable and do not need to be repeated.^(g)Pain assessments will be conducted at 15, 30, and 45 minutes and 1,1.5, 2, 3, 4, 5, 6, 7, 8, 10, and 12 hours after Time 0 and immediatelybefore the first dose of rescue medication. Pain intensity will also beassessed predose. At each assessment time point, the pain intensityassessment will be completed first and the pain relief assessment willbe completed second. Subjects will not be able to compare theirresponses with their previous responses. ^(h)Two stopwatches will bestarted immediately after the subject has swallowed the study drug with8 ounces of water (Time 0). Subjects will record the time to firstperceptible and meaningful pain relief, respectively, by stopping thestopwatches. ^(i)Subjects will complete a global evaluation of studydrug 12 hours after Time 0 or immediately before the first dose ofrescue medication (whichever occurs first). ^(j)Adverse events will bemonitored and recorded from the time of signing of the informed consentform (ICF) until the Follow-up Visit (or Early Termination visit).

A: Screening (Days −28 to −1); B: Day of Surgery (Day 1); C: Preop; D:Postop; E: Predose; F: 0 h; G: 15, 30, 45 min; H: 1, 1.5, 2, 3, 4, 5, 6,7, 8, 10 h; I: 12 h; J: Day 2; K: Follow-up (Day 8±2 days or ET).

Abbreviations: BMI, body mass index; ET, early termination; h, hour;min, minute; preop, preoperative; postop, postoperative; VAS, VisualAnalogue Scale. ^(a) Times listed are relative to dosing with studydrug.^(b) Medical history and concomitant medication use since screeningwill be updated on Day 1 before surgery.^(c) A complete physicalexamination (excluding the genitourinary examination) will be performedat screening. An abbreviated confirmatory physical assessment, includingan examination of the subject's mouth and neck, will be performed at theFollow-Up visit (or Early Termination visit)^(d) Vital signs will berecorded after the subject has been in a sitting position for 5 minutesat the following times: at screening, before surgery, before Time 0, 12hours after Time 0, and/or immediately before the first dose of rescuemedication, and the Follow-up Visit (or Early Termination visit).^(e)Serum pregnancy test at screening and urine pregnancy test beforesurgery on Day 1 (female subjects of childbearing potential only). Testresults must be negative for the subject to continue in the study.^(f)Oral radiographs taken within 1 year before screening will be acceptableand do not need to be repeated.^(g) Pain assessments will be conductedat 15, 30, and 45 minutes and 1, 1.5, 2, 3, 4, 5, 6, 7, 8, 10, and 12hours after Time 0 and immediately before the first dose of rescuemedication. Pain intensity will also be assessed predose. At eachassessment time point, the pain intensity assessment will be completedfirst and the pain relief assessment will be completed second. Subjectswill not be able to compare their responses with their previousresponses.^(h) Two stopwatches will be started immediately after thesubject has swallowed the study drug with 8 ounces of water (Time 0).Subjects will record the time to first perceptible and meaningful painrelief, respectively, by stopping the stopwatches.^(i) Subjects willcomplete a global evaluation of study drug 12 hours after Time 0 orimmediately before the first dose of rescue medication (whichever occursfirst).^(j) Adverse events will be monitored and recorded from the timeof signing of the informed consent form (ICF) until the Follow-up Visit(or Early Termination visit).

The invention claimed is:
 1. A solid oral unit dose of a pharmaceuticalcomposition containing 18 mg of diclofenac acid, wherein the D(90) ofthe diclofenac acid, on a particle volume basis, is less than 5000 nm,and wherein the unit dose, when tested in vitro by USP Apparatus I(Basket) method of U.S. Pharmacopoeia at 100 rpm at 37° C. in 900 ml of0.05% sodium lauryl sulfate in citric acid solution buffered to pH 5.75,has a dissolution rate of diclofenac acid such that 94%, by weight, isreleased by 75 minutes.
 2. The unit dose of claim 1, wherein the D(90)of the diclofenac acid, on a particle volume basis, is less than 3000nm.
 3. The unit dose of claim 2, wherein the unit dose has a dissolutionrate of diclofenac acid such that 94%, by weight, is released by 60minutes.
 4. The unit dose of claim 2, wherein the unit dose has adissolution rate of diclofenac acid such that 94%, by weight, isreleased by 45 minutes.
 5. The unit dose of claim 2, wherein the unitdose has a dissolution rate of diclofenac acid such that 94%, by weight,is released by 30 minutes.
 6. The unit dose of claim 1, wherein theD(90) of the diclofenac acid, on a particle volume basis, is less than2000 nm.
 7. The unit dose of claim 6, wherein the unit dose has adissolution rate of diclofenac acid such that 94%, by weight, isreleased by 60 minutes.
 8. The unit dose of claim 6, wherein the unitdose has a dissolution rate of diclofenac acid such that 94%, by weight,is released by 45 minutes.
 9. The unit dose of claim 6, wherein the unitdose has a dissolution rate of diclofenac acid such that 94%, by weight,is released by 30 minutes.
 10. The unit dose of claim 1, wherein theD(90) of the diclofenac acid, on a particle volume basis, is less than1900 nm.
 11. The unit dose of claim 1, wherein the D(90) of thediclofenac acid, on a particle volume basis, is less than 1800 nm. 12.The unit dose of claim 1, wherein the unit dose has a dissolution rateof diclofenac acid such that 94%, by weight, is released by 60 minutes.13. The unit dose of claim 1, wherein the unit dose has a dissolutionrate of diclofenac acid such that 94%, by weight, is released by 45minutes.
 14. The unit dose of claim 1, wherein the unit dose has adissolution rate of diclofenac acid such that 94%, by weight, isreleased by 30 minutes.
 15. The unit dose of claim 1, wherein a singledose provides perceptible pain relief to a patient suffering from acutepain.
 16. A solid oral unit dose of a pharmaceutical compositioncontaining 35 mg of diclofenac acid, wherein the D(90) of the diclofenacacid, on a particle volume basis, is less than 5000 nm, and wherein theunit dose, when tested in vitro by USP Apparatus I (Basket) method ofU.S. Pharmacopoeia at 100 rpm at 37° C. in 900 ml of 0.05% sodium laurylsulfate in citric acid solution buffered to pH 5.75, has a dissolutionrate of diclofenac acid such that 95%, by weight, is released by 75minutes.
 17. The unit dose of claim 16, wherein the D(90) of thediclofenac acid, on a particle volume basis, is less than 3000 nm. 18.The unit dose of claim 17, wherein the unit dose has a dissolution rateof diclofenac acid such that 95%, by weight, is released by 60 minutes.19. The unit dose of claim 17, wherein the unit dose has a dissolutionrate of diclofenac acid such that 95%, by weight, is released by 45minutes.
 20. The unit dose of claim 17, wherein the unit dose has adissolution rate of diclofenac acid such that 95%, by weight, isreleased by 30 minutes.
 21. The unit dose of claim 16, wherein the D(90)of the diclofenac acid, on a particle volume basis, is less than 2000nm.
 22. The unit dose of claim 21, wherein the unit dose has adissolution rate of diclofenac acid such that 95%, by weight, isreleased by 60 minutes.
 23. The unit dose of claim 21, wherein the unitdose has a dissolution rate of diclofenac acid such that 95%, by weight,is released by 45 minutes.
 24. The unit dose of claim 21, wherein theunit dose has a dissolution rate of diclofenac acid such that 95%, byweight, is released by 30 minutes.
 25. The unit dose of claim 16,wherein the D(90) of the diclofenac acid, on a particle volume basis, isless than 1900 nm.
 26. The unit dose of claim 16, wherein the D(90) ofthe diclofenac acid, on a particle volume basis, is less than 1800 nm.27. The unit dose of claim 16, wherein the unit dose has a dissolutionrate of diclofenac acid such that 95%, by weight, is released by 60minutes.
 28. The unit dose of claim 16, wherein the unit dose has adissolution rate of diclofenac acid such that 95%, by weight, isreleased by 45 minutes.
 29. The unit dose of claim 16, wherein the unitdose has a dissolution rate of diclofenac acid such that 95%, by weight,is released by 30 minutes.
 30. The unit dose of claim 16, wherein asingle dose provides perceptible pain relief to a patient suffering fromacute pain.